Antirheumatic

ABSTRACT

A novel antirheumatic agent comprising as an active ingredient a compound of formula I:                    
     or a pharmaceutically acceptable salt or hydrate thereof.

TECHNICAL FIELD

The present invention relates to novel benzylidene derivatives, and anantirheumatic agent, an osteoclast formation inhibiting agent, anitrogen monoxide production inhibiting agent, and a transcriptionfactor NFκB suppressing agent comprising said benzylidene derivative asan active ingredient. The present invention further relates to a methodfor the prevention and/or treatment of rheumatic diseases, a method forthe inhibition of osteoclast formation, a method for the inhibition ofnitrogen monoxide production, and a method for the suppression oftranscription factor NFκB.

BACKGROUND ART

Rheumatic disease is a systemic inflammatory disease involving afunctional abnormality accompanied by swellings and pains in arthrosisas the primary symptom. The number of the patients in Japan is estimatedas more than about 600 thousands (morbidity: about 0.5% of the Japanesepopulation), and many of these patients are females of middle toadvanced age. Although the development of this disease has not yet beenwell elucidated, it is believed to be a trigger for this disease that animmunological abnormality occurs for some reason, the autoimmuneresponse is accelerated, and an activated macrophage, neutrophil, T-celland the like are infiltrated to inflammatory focus. Then, these cellsreceive an inflammatory stimulus to produce various inflammatorymediators such as IL-1, IL-6, THF-α, nitrogen monoxide (NO) andprostaglandin E₂ (PGE₂). The mediators then induce a cell adhesionfactor, collagenase, protease and the like, which cause swelling andpain and accelerate the destruction of articular cartilage and bone.

According to the recent findings, activation of NFκB due to inflammatorystimulation induces cyclooxygenase-2 (COX-2) and inductive nitrogenmonoxide synthase (i-NOS), and also accelerates an expression ofcytokines. Cytokines and NO produced in the inflammatory focus activateNFκB.

As a result, it is believed that the production of inflammatorymediators is enhanced in the inflammatory focus and the chronicinflammation pathology accompanied by the tissue destruction is thusdeveloped. NO is also associated with the activation of COX-2 andaccelerates production of PGE₂.

Certain benzylidene compounds exhibit an anti-inflammatory effect, andtherefore, they are currently used the treatment of rheumatic diseasesas a nonsteroidal anti-inflammatory drug (NSAID), for the purpose ofsuppressing pains and swellings. However, the use of such nonsteroidalanti-inflammatory drug is a mere symptomatic therapy for the suppressionof pains and swellings due to rheumatic diseases and is not curable forthe disease itself.

On the other hand, antirheumatic agents (disease-modifying antirheumaticdrug) inhibit the progression of rheumatic pathology and they are usedin a radical therapy for the disease. The compounds of the presentinvention have been found to inhibit articular bone destruction, arheumatic pathology, in a rheumatic model animal (NZB/KN) to which anonsteroidal anti-inflammatory drug is not effective. Thus, thecompounds have been found to be effective in a radical therapy ofrheumatic diseases via a different mechanism from that ofanti-inflammatory agents.

Combination use of a disease-modifying antirheumatic drug (DMARD) suchas immunosuppressants or immunomodulators and a nonsteroidalanti-inflammatory drug (NSAID) is often conducted in the treatment ofchronic arthritis such as rheumatoid arthritis. However, these drug havedrawbacks in that the former may often cause serious side effects suchas blood disorder, and the latter may cause gastrointestinal disorderdue to prolonged administration. Accordingly, a drug which possesses theDMARD and NSAID effects but less side effects would be very useful as atherapeutic agent for rheumatoid arthritis.

SUMMARY OF THE INVENTION

The present inventors have found that particular benzylidenederivatives, i.e. the compounds of formula I:

wherein

R¹ is hydrogen, lower alkyl, lower alkoxy, hydroxy lower alkyl, orcarboxy lower alkyl and R⁴ is hydroxy; or

R¹ and R⁴ taken together form —CR⁵R⁶—(CH₂)_(m)—O— or—CR⁵R⁶—(CH₂)_(p)CH(OH)—O—

wherein m is an integer of 1 to 3;

p is an integer of 0 to 2; and

R5 and R6 are each independently hydrogen, lower alkyl, lower alkoxy, orhydroxy lower alkyl;

R² is hydrogen, lower alkyl, lower alkoxy, hydroxy lower alkyl, orcarboxy lower alkyl; and

R³ is hydrogen, lower alkyl, cycloalkyl, lower alkoxy, arylalkyloxy,heteroarylalkyloxy, lower alkylcarbonyl, arylcarbonyl, substituted orunsubstituted carbamoyl or the group represented by the formula:

—(CH₂)_(n)—Q

wherein Q is hydroxy, substituted or unsubstituted amino, aryl,heteroaryl, hydroxycarbonyl or lower alkyloxycarbonyl; and

n is an integer of 0 to 3

 exhibit not only anti-inflammatory effect but also antirheumaticeffect. The present invention has been accomplished on the basis of theabove finding.

The present inventors also have found that the above compounds inhibitenhanced formation of osteoclast caused by PGE₂.

Also, the above compounds have been found to suppress an induciblenitrogen monoxide synthase (i-NOS) to inhibit NO production.

Further, the above compounds have been found to posses suppressiveactivity on a transcription factor NFκB.

Accordingly, the present invention provides novel uses of suchderivatives as an antirheumatic agent, an osteoclast formationinhibiting agent, a nitrogen monoxide production inhibiting agent, and atranscription factor NFκB suppressing agent.

Thus, the present invention provides an antirheumatic agent comprising acompound of formula I or pharmaceutically acceptable salt or hydratethereof as an active ingredient.

The present invention also provides an osteoclast formation inhibitingagent comprising a compound of formula I or pharmaceutically acceptablesalt or hydrate thereof as an active ingredient.

The present invention further provides a nitrogen monoxide productioninhibiting agent comprising a compound of formula I or pharmaceuticallyacceptable salt or hydrate thereof as an active ingredient.

The present invention yet further provides a transcription factor NFκBsuppressing agent comprising a compound of formula I or pharmaceuticallyacceptable salt or hydrate thereof as an active ingredient.

The compounds of the formula I wherein R¹ and R² are both t-butyl, morepreferably R¹ and R² are both t-butyl and R⁴ is hydroxy, more preferablyR¹ and R² are both t-butyl, R³ is lower alkyl and R⁴ is hydroxy, muchmore preferably R¹ and R² are both t-butyl, R³ is ethyl and R⁴ ishydroxy, are preferred antirheumatic agents, osteoclast formationinhibiting agents, nitrogen monoxide production inhibiting agents andtranscription factor NFκB suppressing agents.

The present invention further provides use of the compounds of formula Ifor the manufacture of anti-rheumatic agent.

Also, the present invention provides use of the compounds of formula Ifor the manufacture of osteoclast formation inhibiting agent.

Further, the present invention provides use of the compounds of formulaI for the manufacture of nitrogen monoxide production inhibiting agent.

Yet further, the present invention provides use of the compounds offormula I for the manufacture of transcription factor NFκB suppressingagent.

The present invention further provides a method for the preventionand/or treatment of rheumatoid arthritis which comprises administeringan effective amount of a compound of formula I to mammals in needthereof.

Also, the present invention provides a method for the inhibition ofosteoclast formation which comprises administering an effective amountof a compound of formula I to mammals in need thereof.

Further, the present invention provides a method for the inhibition ofnitrogen monoxide production which comprises administering an effectiveamount of a compound of formula I to mammals in need thereof.

Yet further, the present invention provides a method for the suppressionof transcription factor NFκB which comprises administering an effectiveamount of a compound of formula I to mammals in need thereof.

Among the compounds represented by the above formula I, some compoundshave been disclosed as inhibiting agent against PGE₂ production inJapanese Patent Publication No. 211819/1994 (published on Aug. 2, 1994).

The present invention further provides novel compounds of formula I′:

wherein

R^(1′) is hydroxy lower alkyl or carboxy lower alkyl and R^(4′) ishydroxy; or

R^(1′) and R_(4′) taken together form —CR⁵R⁶—(CH₂)_(m)—O— or—CR⁵R⁶—(CH₂)_(p)CH(OH)—O—

wherein m is an integer of 1 to 3;

p is an integer of 0 to 2; and

R5 and R6 are each independently hydrogen, lower

alkyl, lower alkoxy, or hydroxy lower alkyl;

R^(2′) is hydrogen, lower alkyl, lower alkoxy, hydroxy lower alkyl, orcarboxy lower alkyl; and

R^(3′) is hydrogen, lower alkyl, cycloalkyl, lower alkoxy, arylalkyloxy,heteroarylalkyloxy, lower alkylcarbonyl, arylcarbonyl, substituted orunsubstituted carbamoyl or the group represented by the formula:

—(CH₂)_(n)—Q

wherein Q is hydroxy, substituted or unsubstituted amino, aryl,heteroaryl, hydroxycarbonyl or lower alkyloxycarbonyl; and

n is an integer of 0 to 3

or pharmaceutically acceptable salt or hydrate thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the inhibitory effect of the compound of theinvention against adjuvant articular swelling. Values in the graph aremean of the eight independent experiments, and significance test wasperformed according to Dunnett's t-test (*p<0.05, **p<0.01: vs. vehiclegroup).

FIG. 2 is a X-ray observation score for bone destruction in arthrosis.

FIG. 3 is a graph showing the inhibitory effect against the destructionof articular bone in adjuvant arthritis. Values in the graph are meanstandard error of eight independent experiments, and significance testwas performed according to Dunnett's t-test (*p<0.05, **p<0.01: vs.vehicle group).

FIG. 4 is a graph showing the repressive effect on the IL-6 level inarticular tissue. Nor: normal rat, Veh: vehicle, Buc: Bucillamine, CsA:Cyclosporin A. Values in the graph are mean±standard error of sevenindependent experiments, and significance test was performed accordingto Dunnett's t-test (##p<0.01: vs. normal group, **p<0.01: vs. vehiclegroup).

FIG. 5 is a graph showing the inhibitory effect against bone destructionin NZB/KN mouse. Veh: vehicle, CsA: Cyclosporin A, Ind: Indomethacin,Ten: Tenidap. N refers to the number of animals tested. Values in thegraph are mean +standard error, and significance test was performedaccording to Dunnett's t-test (*p<0.05, **p<0.01: vs. vehicle group).

FIG. 6 is a graph showing the repressive effect on MRL/1 mouse bloodacute phase protein SAP. Veh: vehicle, Buc: Bucillamine, Lob:Lobenzarit, Ten: Tenidap, Ind: Indomethacin, and Pred: Predonisolone.Values in the graph are mean±standard error, and significance test wasperformed according to Dunnett's t-test (*p<0.05, **p<0.01: vs. vehiclegroup).

FIG. 7 is a graph showing the repressive effect on suppressive effect onblood NOx level in adjuvant arthritis rat. Nor: normal rat, Veh:vehicle, Ind: Indomethacin, and Dex: Dexamethazon. Values in the graphare mean±standard error of eight independent experiments, andsignificance test was performed according to Dunnett's t-test (##p<0.01:vs. normal group, *p<0.05, **p<0.01: vs. vehicle group).

DETAILED DESCRIPTION

As is apparent from the above formula, the compounds of the invention offormula I can exist in the stereostructures of (E)-type. Accordingly,unless otherwise noted, the compound I described in this specificationshould be construed to include (E)-isomer.

For purposes of the present invention, as disclosed and claimed herein,the following terms are defined as below.

The term “lower alkyl” means straight or branched C₁-C₈ alkyl andexamples thereof include methyl, ethyl, n-propyl, i-propyl n-butyl,i-butyl, s-butyl, t-butyl, n-pentyl, i-pentyl, neopentyl, s-pentyl,t-pentyl, n-hexyl, neohexyl, i-hexyl, s-hexyl, t-hexyl, heptyl andoctyl. Among them, a straight or branched C₁-C₄ alkyl is preferred. Themost preferred one is methyl or ethyl.

The term “lower alkoxy” means straight or branched C₁-C₆ alkyloxy andexamples thereof include methoxy, ethoxy, n-propoxy, i-propoxy,n-butoxy, i-butoxy, s-butoxy, t-butoxy, n-pentyloxy, i-pentyloxy,neopentyloxy, s-pentyloxy, t-pentyloxy, n-hexyloxy, neohexyloxy,i-hexyloxy, s-hexyloxy and t-hexyloxy and the like. Among them, a C₁-C₃alkoxy group is preferred. The most preferred one is methoxy.

The term “hydroxy lower alkyl” means a group formed by substituting thelower alkyl group defined above with hydroxy group(s). Specific examplesthereof include hydroxymethyl, hydroxyethyl, hydroxypropyl,hydroxybutyl, 1,1-dimethyl-2-hydroxyethyl and the like.

The term “carboxy lower alkyl” means a group formed by substituting thelower alkyl group defined above with carboxy group(s). Examples thereofinclude carboxymethyl, carboxyethyl, carboxypropyl, carboxybutyl,1,1-dimethyl-2-carboxyethyl and the like.

The term “cycloalkyl” means C₃-C₇ cycloalkyl and examples thereofinclude cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyland the like. Among them, C₃-C₅ cycloalkyl is preferred.

The term “aryl” means unsubstituted or substituted phenyl or naphthyland may be substituted by one or more substituents including halogen,lower alkoxy, lower alkyl, nitro and the like. Examples of aryl includephenyl, 4-chlorophenyl, 4-methoxyphenyl, 4-nitrophenyl,3,4-dichlorophenyl, 3,4-dimethoxyphenyl, 3,4-dinitrophenyl, 1-naphtyl,2-naphtyl and the like.

The term “arylalkyloxy” means a group formed by substituting a loweralkoxy group with an aryl group(s) as defined above and examples thereofinclude benzyloxy, 4-chlorobenzyloxy, 4-methoxybenzyloxy,3,4-dichlorobenzyloxy, 3,4-dimethoxybenzyloxy, 4-nitrobenzyloxy,2-phenylethyloxy, 2-(4-chlorophenyl)ethyloxy,2-(4-methoxyphenyl)ethyloxy, 1-naphtylmethyloxy, 2-naphtylmethyloxy andthe like. Among them, benzyloxy is preferred.

The term “heteroaryl” means a group containing 1-4 hetero atoms andexamples thereof include pyridyl, thiazolyl, isothiazolyl, oxazolyl,isoxazolyl, imidazolyl, triazolyl and tetrazolyl. For purposes of thepresent invention, pyridyl, thiazolyl, oxazolyl and imidazolyl arepreferred and pyridyl is most preferred.

The term “heteroarylalkyloxy” means a group formed by substituting analkoxy group with a heteroaryl group(s) as defined above and examplesthereof include 2-pyridylmethyloxy, 3-pyridylmethyloxy,4-pyridylmethyloxy, 2-imidazolylmethyloxy, 4-imidazolylmethyloxy,2-thiazolylmethyloxy, 4-thiazolylmethyloxy and the like.

Examples of “lower alkylcarbonyl” include acetyl, propionyl, butyryl,valeroyl, hexanoyl, heptanoyl and octanoyl.

Examples of “arylcarbonyl” include benzoyl, 4-chlorobenzoyl,4-methoxybenzoyl, 4-nitrobenzoyl, 3,4-dichlorobenzoyl,3,4-dimethoxybenzoyl, 3,4-dinitrobenzoyl, 1-naphthoyl, 2-naphthoyl andthe like.

For the term “substituted or unsubstituted carbamoyl”, examples of thesubstituent therefor include lower alkyl, lower alkoxy, hydroxy,cycloalkyl, arylalkyl, alkoxyalkyl, alkylcarbonyl, arylcarbonyl,cycloalkyloxy and arylalkyloxy, and one or more of these substituentsmay be substituted at the nitrogen atom. Among them, lower alkyl, loweralkoxy and hydroxy are preferred. Examples of substituted carbamoylinclude N-methylcarbamoyl, N,N-dimethylcarbamoyl, N-hydroxycarbamoyl,N-methyl-N-hydroxycarbamoyl, N-methoxycarbamoyl,N-methoxy-N-methylcarbamoyl, N-ethylcarbamoyl, N,N-diethylcarbamoyl,N-ethyl-N-hydroxycarbamoyl, N-propylcarbamoyl, N,N-dipropylcarbamoyl andN-propyl-N-hydroxycarbamoyl.

Examples of “halogen” include fluorine, chlorine, bromine and iodine.

Examples of “lower alkyloxycarbonyl” include methoxycarbonyl,ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl,isobutoxycarbonyl, tert-butoxycarbonyl and the like.

The term “substituted amino” means mono- or di-substituted amino, andexamples of substituent include lower alkyl and arylalkyl as definedabove.

The term “pharmaceutically acceptable salt or hydrate” refers to saltsor hydrates of the compounds represented by the formula I which arenon-toxic to a living thing. Typical pharmaceutically acceptable saltsinclude inorganic or organic salts formed by the reaction of a compoundof formula I with an inorganic or organic acid or base. Such salts areknown as acid or base addition salts.

Examples of commonly used acids to form acid addition salts includeinorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodicacid, sulfuric acid, phosphoric acid and the like, and organic acidssuch as p-toluene sulfonic acid, methanesulfonic acid, oxalic acid,p-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid,benzoic acid, acetic acid and the like.

Example of commonly used bases to form base addition salts includeinorganic bases such as ammonium hydroxide, ammonium carbonate, ammoniumhydrogen carbonate, alkaline metal or alkaline earth metal hydroxides orcarbonates and the like, and organic bases such as aliphatic amines,primary, secondary and tertiary amines, and aliphatic diamines and thelike.

In general, the above pharmaceutically acceptable acid or base additionsalts can be obtained by the reaction of a compound of formula I withequimolar or excess amount of an acid or base.

The compound of the invention can be essentially prepared by the processdescribed in the Japanese Patent Publication No. 211819/1994, thedisclosure of which is herein incorporated by reference. However, thepresent invention is by no means limited by the process per se. Oneprocess for the production of the compounds of the invention isexemplified in the following Preparations.

In the above formulae, R¹, R², and R³ are as defined above and R⁷ ishydrogen or a hydroxy-protecting group.

In the case that R⁷ of the compound 6 is a hydroxy-protecting group,examples of the protecting group include methoxymethyl, methoxyethyl,trimethylsilyl, tert-butyldimethylsilyl. R⁷ is preferably ahydroxy-protecting group, particularly methoxymethyl group.

One of the starting compounds for the above reaction, thesulfur-containing compound 4 can be prepared, for example, according toa reaction scheme below.

In the above formulae, R³ is as defined above.

3-Chloropropylsulfonyl chloride 1 is reacted with amine 2 to yieldsulfonamide intermediate 3.

The reaction is carried out in the presence of base (A), if necessary,in a solvent such as ether, chloroform, methylene chloride,dichloroethane, tetrahydrofuran, dimethoxyethane, diethoxyethane,benzene, toluene, xylene, ethyl acetate, methyl acetate and the like,which solvent may contain water. The amine (R³NH₂) may be in the form ofhydrochloride salt.

The base (A) used in the case of necessity includes alkali metal basessuch as lithium hydroxide, sodium hydroxide, potassium hydroxide, sodiumcarbonate, potassium carbonate, sodium bicarbonate and potassiumhydrogencarbonate and the like, and organic bases such as pyridine,4-N,N-dimethylaminopyridine (DMAP), triethylamine, diisobutylethylamine, 1,8-diazabicyclo[5,4,0]undec-7-ene (DBU),1,4-diazabicyclo[2,2,2]octane (DABCO) and the like. When an alkali metalbase is used, it is preferable to add a phase transfer catalyst, ifnecessary. Examples of preferred phase transfer catalysts are quaternaryammonium salts such as N-benzyltrimethylammonium salts,tetrabutylammonium salts and the like.

The reaction for converting sulfonamide intermediate 3 intosulfur-containing heterocyclic compound 4 can be carried out in thepresence of a base (B) in a solvent as described above, while anhydroussolvents such as dimethyl sulfoxide, dimethylformamide and the like mayalso be used and rather preferable. Sodium hydride and lithium hydridecan be used as a base B as well as those described above.

Alternatively, sulfur-containing compound 4 can be prepared directlyfrom compound 1 without separation of sulfonamide intermediate 3. Inthis case, the reaction of compound 1 with amine 2 is carried out in asuitable solvent in the presence of two equivalents of a base. Thesolvent and the base may be selected from those exemplified above but itis particularly preferable to use sodium hydride as a base anddimethylformamide as a solvent.

Alternatively, the desired sulfur-containing compound 4 can also beobtained from commercially available γ-sultone 5. (See “Preparation”).Briefly, compound 5 is allowed to react with amine (R³NH₂) and theresultant product is then treated with a dehydrating agent. The reactioncan be carried out without solvent but may be conducted in a solventdescribed above, if necessary. Commonly used reagent such as phosphorusoxychloride, thionyl chloride, phosphorus pentachloride, phosphoruspentoxide and the like may be used as a dehydrating agent, andphosphorus oxychloride is particularly preferable.

The aldol reaction between compounds 6 and 4 obtained above is carriedout in the presence of a base (C) in a suitable solvent. Examples ofbase (C) include organic lithium salts such as n-butyllithium,sec-butyllithium, tert-butyllithium, phenyllithium, lithiumdiisopropylamide, lithium diethylamide, lithium hexamethyldisilazane andthe like, and alkali metal bases such as sodium hydride and potassiumtert-butoxide and the like. Particularly, lithium diisopropylamide orlithium hexamethyldisilazane is preferable.

Examples of reaction solvents include ether solvents such as diethylether, tetrahydrofuran (THF), dimethoxyethane, diethoxyethane and thelike or hydrocarbon solvents such as n-hexane, cyclohexane and the like.The reaction is preferably conducted in the presence of a reagent thatserves as a ligand of lithium metal, for exampletetramethylethylenediamine, hexamethylphosphoramide and the like, ifnecessary.

The reaction is carried out at temperature ranging from −80° C. to +50°C. with preference in lower temperature range.

Aldol adduct 7 is converted to compound I in the presence of an acid.Examples of acids include organic acids such as trifluoroacetic acid,p-toluenesulfonic acid, camphorsulfonic acid and the like and inorganicacids such as sulfuric acid, hydrochloric acid and the like. Further,ordinary dehydrating agents such as thionyl chloride, methanesulfonylchloride, aluminium chloride, phosphorus oxychloride, phosphoruspentachloride and the like can be used. Preferably, the reaction iscarried out with heating in an aromatic hydrocarbon such as benzene,toluene, xylene and the like, a halogenated hydrocarbon such aschloroform, dichloromethane, dichloroethane and the like, or an ethersolvent such as tetrahydrofuran, dimethoxyethane, diethoxyethane and thelike.

Base (C) in the above formula is as defined above. Y means anN-protecting group such as tert-butoxycarbonyl, benzyloxycarbonyl,benzyl, 4-methoxybenzyl, 3,4-dimethoxybenzyl, 4-nitrobenzyl and thelike. The reaction conditions of the aldol reaction are similar to thosedescribed in the reaction scheme I above. Dehydrating and deprotectingreagents used in the conversion of aldol adduct 7′ to compounds 10include p-toluenesulfonic acid and trifluoroacetic acid, aluminiumchloride, titanium tetrachloride and the like. The conditions such asreaction solvent, temperature and the like are similar to thosedescribed in reaction scheme I. Compound 8 is deprotected to obtaincompound 10, which is the compound of formula I wherein R³ is hydrogen.

In this reaction, a desired substituent R³ is added to compound 10obtained in the above reaction scheme II, which corresponds to thecompound of formula I wherein R³ is hydrogen in the formula I, to yieldvarious derivatives. Base (D) to be used when R³—X is an alkylatingagent includes alkali metal salts such as sodium hydroxide, potassiumhydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate,potassium hydrogen carbonate, lithium hydroxide and the like, or organicbases such as pyridine, triethylamine, diisopropylethylamine and thelike. The alkylation is preferably carried out using sodium hydroxide orpotassium carbonate in the presence of an appropriate quaternaryammonium salt as a phase transfer catalyst.

In the case where R³X is an acylating agent, an organic base such aspyridine, 4-dimethylaminopyridine, triethylamine, diisopropylethylamineor the like is preferably used as base (D).

When R³X is a carbamoylating agent or alkoxycarbonylating agent, anorganic lithium base such as n-butyllithium, lithiumhexamethyldisilazane, lithium diisopropylamide or the like is preferablyutilized as base (D). The present invention is not limited to the use ofthese bases, and organic bases such as pyridine, triethylamine,diisopropylethylamine and the like or the alkali metal bases describedabove are also useable.

Compounds 12, which corresponds to the compound of formula I wherein R³is OH, is obtained by de-benzylation of compounds 11. The de-benzylationis carried out using a deprotecting agent. The deprotection is conductedby hydrogenation in the presence of palladium on carbon or platinumoxide as catalyst, or by using a Lewis acid such as aluminium chloride,titanium tetrachloride or the like along with anisole,2,6-di-tert-butylphenol and the like, if necessary.

Besides halogenated hydrocarbons such as dichloromethane, chloroform,dichloroethane and the like, nitromethane, benzene, toluene, xylene andthe like can also be used as a reaction solvent.

The processes in the above schemes I, II and III are generallyapplicable to the production of compounds I of the invention, and theyare specifically illustrated in Preparations 1-43 below.

The compound I of the present invention can be orally or parenterallyadministered as an anti-inflammatory agent. In the case of oraladministration, a compound of the present invention may be formulatedinto ordinary formulations in the form of solid formulations such astablets, powders, granules, capsules and the like; solutions; oilysuspensions; or liquid formulations such as syrups, elixirs and thelike. In the case of parenteral administration, a compound of thepresent invention may be formulated into an aqueous or oily suspensionfor injection or an external preparation. In preparing the formulations,conventional excipients, binders, lubricants, aqueous solvents, oilysolvents, emulsifiers, suspending agents or the like may be used, andother additives, such as preservatives, stabilizers or the like may alsobe included.

Although appropriate daily dosage of the compound of the inventionvaries depending upon the administration route, age, body weight andconditions of a particular patient, and the kind of disease to betreated. In general, however, in the case of adult patients, it may bebetween 10-500 mg, preferably 50-100 mg for oral administration, and1-250 mg, preferably 5-10 mg for parenteral administration, in 1-5divisions.

The following Preparations and Examples are provided to furtherillustrate the present invention and are not to be construed as limitingthereof. The abbreviations used in the Preparations and Examples havethe following meanings:

LDA: lithium diisopropylamide;

MOM: methoxymethyl;

p-TsOH: p-toluenesulfonic acid;

THF: tetrahydrofuran;

DMF: N,N-Dimethylformamide;

HMPA: hexamethylphosphoramide;

LiHMDS: lithium hexamethyldisilazane;

DBU: 1,8-diazabicyclo[5,4,0]undec-7-ene; and

DIBAL: diisobutylalminium hydride.

Preparation 1 (R³=Et) N-Ethyl-1,2-isothiazolidine-1,1-dioxide (4a)

To a solution of 3-chloropropylsulfonyl chloride 1 (6.1 g, 34.5mmol) inether (25 ml) was dropwise added ethylamine (a 70% aqueous solution, 4.4g, 68.3 mmol) with stirring and ice-cooling over 15 minutes, and theresultant mixture was stirred for one hour at room temperature. Thereaction mixture was concentrated in vacuo, Benzene (100 ml) was addedto the residue, and the solvent was removed in vacuo. To the residue wasadded ether (150 ml) and filtered to remove the insoluble material. Thefiltrate was distilled in vacuo to remove ether and 6.96 g (yield, about100%) of crude N-ethyl-3-chloro propylsulfonamide (intermediate 3a) wasobtained as colorless crystals (m.p.=30-32° C.). To a solution of thisintermediate 3a (6.96 g, 34.5 mmol) in THF (50 ml) was slowly addedsodium hydride (60% in oil, 1.52 g, 38.0 mmol) with stirring underice-cooling over 15 minutes. The reaction mixture was stirred foranother 30 minutes at room temperature. After the addition of ether (50ml), the mixture was filtered to remove insoluble material and thesolvent was distilled in vacuo to give 4.93 g (96%) of the desiredcompound 4a as a pale yellow oil. IR (CHCl₃)cm⁻¹: 3018, 2976, 2868,1452, 1306, 1220, 1179, 1129, 1015; NMR (CDCl₃) δ: 1.24 (3H, t, J=7.4Hz, CH₃), 2.28-2.42 (2H, m, CH₂), 3.10 (2H, q, J=7.4 Hz, CH₂), 3.15 (2H,t, J=7.6 Hz, CH₂), 3.22-3.29 (2H, m, CH₂).

Preparation 2 (R³=Me) N-Methyl-1,2-isothiazolidine-1,1-dioxide (4b)

3-Chloropropylsulfonyl chloride 1 (16.8 g, 94.9 mmol), ethylaminehydrochloride (13.5 g, 200 mmol), and potassium carbonate (27.6 g, 200mmol) were sequentially added to ethyl acetate (500 ml). After theaddition of N-benzyltrimethylammonium chloride (about 200 mg) to themixture, the resultant mixture was stirred for 2 hours at roomtemperature and dried over anhydrous sodium sulfate. The mixture wasfiltered through a small amount of silica² gel and the filtrate wasconcentrated in vacuo to give 12 g (74%) of crudeN-methyl-3-chloropropylsulfonamide (intermediate 3b) as a pale yellowoil.

To a solution of said intermediate 3b (11.79 g, 68.69 mmol) in benzene(300 ml) was added DBU (10.79 ml, 72.12 mmol), and the resultant mixturewas stirred for 24 hours at room temperature and filtered through asmall amount of silica gel. The solvent was removed to give 7.0 g (75%)of the desired compound 4b as a colorless solid (m.p.=36-40° C.). IR(CHCl₃)cm⁻¹: 3016, 1451, 1307, 1218, 1187, 1127; NMR (CDCl₃) δ:2.27-2.42 (2H, m, CH₂), 2.69 (3H, s, CH₃), 3.11-3.20 (2H, m, CH₂), 3.22(2H, t, J=6.0 Hz, CH₂).

Preparation 3 (R³=CH₂CH(CH₃)₂)N-Isobutyl-1,2-isothiazolidine-1,1-dioxide (4c)

3-Chloropropylsulfonyl chloride 1 (7.08 g, 40 mmol), isobutylamine (7.3g, 100 mmol) and sodium bicarbonate (3.36 g, 40 mmol) were sequentiallyadded to a mixture of ethyl acetate (200 ml) and water (20 ml). To themixture was added N-benzyltrimethylammonium chloride (about 100 mg), andthe resultant mixture was stirred for 3 hours at room temperature andthen treated in a manner described in Preparation 2 to give 8.19 g (96%)of crude N-isobutyl-3-chloropropylsulfonamide (intermediate 3c) ascolorless crystals (m.p.=68-69° C.).

To a solution of said intermediate 3c (4.27 g, 20 mmol) in benzene (60ml) was added DBU (3.3 ml, 22 mmol), and the reaction mixture wastreated in a manner as described in Preparation 2 to give 3.37 g (95%)of the desired compound 4c as a colorless oil. IR (CHCl₃)cm⁻¹: 3016,2956, 1465, 1304, 1226, 1131, 1024; NMR (CDCl₃) δ: 0.95 (6H, d, J=6.6Hz, (CH₃)₂), 1.75-1.96 (1H, m, CH), 2.27-2.42 (2H, m, CH₂), 2.80 (2H, d,J=7.4 Hz, CH₂), 3.10-3.19 (2H, m, CH₂), 3.24 (2H, t, J=6.8 Hz, CH₂).

Preparation 4 (R³=Cyclopropyl)N-Cyclopropyl-1,2-isothiazolidine-1,1-dioxide (4d)

3-chloropropylsulfonyl chloride 1 (7.08 g, 40 mmol), cyclopropylamine(6.0 g, 105 mmol) and sodium bicarbonate (3.7 g, 44 mmol) were treatedin a mixture of ether (200 ml) and water (10 ml) in a manner asdescribed in Preparation 3 to give 8.0 g (about 100%) of crudeN-cyclopropyl-3-chloropropylsulfonamide (intermediate 3d) as crystals(m.p.=48-49.5° C.).

Said intermediate 3d (1.98 g, 10 mmol) was reacted with DBU (1.65 ml, 11mmol) in benzene (30 ml) in a manner as described in Preparation 2 togive 1.40 g (87%) of the desired compound Ad as a pale yellow oil. IR(CHCl₃)cm⁻¹: 3016, 1309, 1221, 1140, 1026; NMR (CDCl₃) δ: 0.60-0.85 (4H,m, cyclopropyl), 2.20-2.40 (2H, m, CH₂), 3.15-3.25 (3H, m, CH₂+CH), 3.32(2H, t, J=6.6 Hz, CH₂).

Preparation 5 (R³=—CH₂CH₂CH₃) N-n-Propyl-1,2-isothiazolidine-1,1-dioxide(4e)

3-chloropropylsulfonyl chloride 1 (7.08 g, 40 mmol), n-propylamine (5.90g, 100 mmol), potassium carbonate (5.52 g, 40 mmol) and a small amountof N-benzyltrimethylammonium chloride (about 100 mg) were stirred in amixture of ether (200 ml) and water (20 ml) for 3 hours, the reactionmixture was then treated in a manner as described in Preparation 2 togive 8.0 g (about 100%) of crude N-n-propyl-3-chloropropylsulfonamide(intermediate 3e) as crystals (m.p.=47.5-48° C.).

Said intermediate 3e (2.0 g, 10 mmol) was reacted with DUB (1.65 ml, 11mmol) in benzene (30 ml) in a manner as described in Preparation 2 togive 1.41 g (86%) of the desired compound 4e as a pale yellow tocolorless oil. IR (CHCl₃)cm⁻¹: 3018, 2962, 2868, 1304, 1224, 1130, 1019;NMR (CDCl₃) δ: 0.96 (3H, t, J=7 Hz, CH₃), 1.52-1.72 (2H, m, CH₂),2.28-2.42 (2H, m, CH₂), 2.94-3.04 (2H, m, CH₂), 3.10-3.20 (2H, m, CH₂),3.25 (2H, t, J=6.7 Hz, CH₂).

Preparation 6 (R³=OCH₃) N-Methoxy-1,2-isothiazolidine-1,1-dioxide (4f)

3-Chloropropylsulfonyl chloride 1 (7.08 g, 40 mmol),O-methylhydroxylamine hydrochloride (3.76 g, 40 mmol), and potassiumcarbonate (5.80 g, 42 mmol) were reacted in a manner as described inPreparation 5 to give 7.02 g (94%) of crudeN-methoxy-3-chloropropylsulfonamide (intermediate 3f) as a colorless topale yellow oil.

The intermediate 3f (6.25 g, 33.3 mmol) was reacted with sodium hydride(60% in oil, 1.47 g, 36.7 mmol) in a manner as described in Preparation1 to give 3.70 g (73%) of the desired compound 4f as a colorless oil. IR(CHCl₃)cm⁻¹: 3022, 1355, 1249, 1222, 1165, 1138, 1035, 1011; NMR (CDCl₃)δ: 2.37-2.50 (2H, m, CH₂), 3.20-3.14 (2H, m, CH₂), 3.50 (2H, t, J=7.0Hz, CH₂), 3.81 (3H, s, OCH₃).

Preparation 7 (R³=OCH₂C₆H₅) N-Benzyloxy-1,2-isothiazolidine-1,1-dioxide(4g)

3-Chloropropylsulfonyl chloride 1 (30.28 g, 0.17 mol),O-benzylhydroxylamine hydrochloride (27.3 g, 0.17 mol), potassiumcarbonate (50 g, 0.36 mol) and tetrabutylammonium sulfate (about 500 mg)are reacted in a mixture of ether (1L) and water (100 ml) for 24 hoursat room temperature and the reaction mixture was extracted with ethylacetate. The extract was subjected to column chromatography on silicagel. From the fraction eluted with a mixture of ethyl acetate/n-hexane(1:4), 18.4 g (41%) of crude N-benzyloxy-3-chloropropylsulfonamide(intermediate 3g) was obtained as a pale yellow oil.

To a solution of the above intermediate 3g (18.4 g, 69.9 mmol) in THF(150 ml) was added sodium hydride (60% in oil, 2.94 g, 73.4 mmol) andthe reaction was carried out in a manner as described in Preparation 1.The product was subjected to column chromatography on silica gel. Fromthe fraction eluted with a mixture of ethyl acetate/n-hexane (1:5),10.75 g (68%) of the desired compound 4g was obtained as a colorlesscrystal. M.p.=52-54° C. IR (CHC_(l) ₃)cm⁻¹: 3022, 2956, 1453, 1354,1165, 1140, 1081, 1000; NMR(CDCl₃) δ: 2.30-2.48 (2H, m, CH₂), 3.04-3.14(2H, m, CH₂), 3.45 (2H, t, J=6.9 Hz, CH₂), 5.00 (2H, s, OCH₂), 7.30-7.45(5H, m, C₆H₅).

Preparation 8 (R³=4-Methoxybenzyl)N-(4-Methoxybenzyl)-1,2-isothiazolidine-1,1-dioxide (4h)

3-Chloropropylsulfonyl chloride 1 (17.7 g, 0.1 mol),p-methoxybenzylamine (15.0 g, 0.11 mol), and sodium bicarbonate (8.4 g,0.1 mol) were reacted in a mixture of ethyl acetate (400 ml) and water(40 ml) in a manner as described in Preparation 3 to give 19.1 g (69%)of crude N-(4-methoxybenzyl)-3-chloropropylsulfonamide (intermediate 3h)as colorless crystals. M.p.=78-80° C.

The above intermediate 3h (11.11 g, 40 mmol) was reacted with DBU (6.6ml, 40 mmol) in benzene (150 ml). The resultant mixture was treated in amanner as described in Preparation 2 to give 8.89 g (92%) of the desiredcompound 4h as crystals. M.p.=48-51° C. IR (CHCl₃)cm⁻¹: 3016, 1612,1511, 1304, 1245, 1136, 1034. NMR (CDCl₃) δ: 2.20-2.38 (2H, m, CH₂),3.09 (2H, t, J=6.8 Hz, CH₂), 3.14-3.24 (2H, m, CH₂), 3.81 (3H, s, OCH₃),4.12 (2H, s, CH₂), 6.84-6.94 (2H, m, CH₂), 7.22-7.32 (4H, m,4×aromatic-H).

Preparation 9 (R³=3,4-Dimethoxybenzyl)N-(3,4-Dimethoxybenzyl)-1,2-isothiazolidine-1,1-dioxide (4i)

3-Chloropropylsulfonyl chloride 1 (8.85 g, 50 mmol),3,4-dimethoxybenzylamine (9.0 ml, 60 mmol) and potassium carbonate (4.14g, 30 mmol) were treated in a manner as described in Preparation 2 togive 14.5 g (94%) of crudeN-(3,4-dimethoxybenzyl)-3-chloropropylsulfonamide (intermediate 3I).From the intermediate 3i, the desired compound 4i (yield: 69%) wasyielded in a manner as described in Preparation 1. IR (CHCl₃)cm⁻¹: 3018,1516, 1307, 1262, 1225, 1155, 1138, 1027. NMR (CDCl₃) δ: 2.22-2.38 (2H,m, CH₂), 3.11 (2H, t, J=6.7 Hz, CH₂), 3.16-3.25 (2H, m, CH₂), 3.88 (3H,s, OCH₃), 3.89 (3H, s, OCH₃), 4.12 (2H, s, CH₂), 6.79-6.91 (3H, m,3×aromatic-H).

Preparation 10 (R³=C₆H₅) N-Phenyl-1,2-isothiazolidine-1,1-dioxide (4j)

3-Chloropropylsulfonyl chloride 1 (1.456 g, 8.23 mmol) was dropwiseadded to a solution of aniline (0.5 ml, 8.23 mmol) in pyridine (5 ml)with cooling at −20° C. to −30° C. over about 5 minutes. After thecompletion of the addition, the reaction mixture was stirred for another45 minutes at room temperature. The reaction mixture was concentrated invacuo and the residue was subjected to column chromatography on silicagel. From the fraction eluted with a mixture of ethyl acetate/n-hexane(1:2), 1.683 g (88%) of N-phenyl-3-chloropropylsulfonamide (intermediate3j) was obtained as a yellow oil. From the intermediate 3j, the desiredcompound 4j (yield: 57%) was yielded as a pale yellow solid in a manneras described in Preparation 1. IR (CHCl₃)cm⁻¹: 3020, 1598, 1495, 1315,1139. NMR (CDCl₃) δ: 2.46-2.60 (2H, m, CH₂), 3.34-3.42 (2H, m, CH₂),3.78 (2H, t, J=6.6 Hz, CH₂), 7.10-7.40 (5H, m, C₆H₅).

Preparation 11 (R³=4-Chlorophenyl)N-(4-Chlorophenyl)-1,2-isothiazolidine-1,1-dioxide (4k)

According to a similar method to that of Preparation 10,3-chloropropylsulfonyl chloride was reacted with 4-chloroaniline inpyridine to give N-(4-chlorophenyl)-3-chloropropylsulfonamide(intermediate 3k) (yield 93%). From the intermediate 3k, the desiredcompound 4k (yield: 68%) was yielded as colorless crystals(m.p.=110.5-111.5° C.) in a manner as described in Preparation 1. IR(KBr)cm⁻¹: 3010, 2960, 1595, 1493, 1300, 1267, 1131. NMR (CDCl₃) δ:2.47-2.61 (2H, m, CH₂), 3.35-3.43 (2H, m, CH₂), 3.76 (2H, t, J=6.4 Hz,CH₂), 7.16-7.36 (4H, m, 4×aromatic-H).

Preparation 12 (R³=2-Pyridyl)N-(2-Pyridyl)-1,2-isothiazolidine-1,1-dioXide (41)

According to a similar method to that of Preparation 10,3-chloropropylsulfonyl chloride was reacted with 2-aminopyridine to giveN-(2-pyridyl)-3-chloropropylsulfonamide (intermediate 3l) as a paleyellow solid (yield 54%). To a solution of this intermediate 3l (2.138g, 9.11 mmol) in DMF (30 ml) was added sodium hydride (60% in oil, 401mg, 10 mmol) under ice-cooling. The resultant mixture was stirred for 30minutes at 85° C. and the solvent was removed in vacuo. The residue wassubjected to column chromatography on silica gel. From the fractioneluted with a mixture of ethyl acetate/n-hexane (1:1), 1.806 g (100%) ofthe desired compound 4l was obtained as a yellow solid. IR (CHCl₃)cm⁻¹:3022, 1592, 1473, 1434, 1139. NMR (CDCl₃) δ: 2.47-2.60 (2H, m, CH₂),3.43 (2H, t, J=7.5 Hz, CH₂), 4.05 (2H, t, J=6.6 Hz, CH₂), 6.88-7.02 (1H,m, CH), 7.26-7.35 (1H, m, CH), 7.58-7.70 (1H, m, CH), 8.33 (1H, d, J=4.4Hz, CH).

Preparation 13 (R³=3-Pyridyl)N-(3-Pyridyl)-1,2-isothizolidine-1,1-dioxide (4m)

According to a similar method to that of Preparation 10,3-chloropropylsulfonyl chloride 1 (7.28 g, 41.1 mmol) was reacted with3-aminopyridine (4.6 g, 49.3 mmol) in pyridine (15 ml) to give 4.50 g(46%) of crude N-(3-pyridyl)-3-chloropropylsulfonamide (intermediate 3m)as a colorless solid.

The intermediate 3m (232 mg, 0.988 mmol) was treated with sodium hydride(60% in oil, 43.5 mg, 1.09 mmol) in DMF (5 ml) in a similar manner asdescribed in Preparation 12 to give 190 mg (97%) of the desired compound4m as a colorless solid. IR (CHCl₃)cm⁻¹: 3022, 2960, 1590, 1484, 1428,1319, 1142. NMR (CDCl₃) δ: 2.53-2.67 (2H, m, CH₂), 3.38-3.45 (2H, m,CH₂), 3.83 (2H, t, J=6.6 Hz, CH₂), 7.28-7.36 (1H, m, CH ), 7.73-7.79(1H, m, CH), 8.41 (1H, d, J=4.6 Hz, CH), 8.46 (1H, d, J=2.4 Hz, CH).

Preparation 14 (R³=4-Pyridyl)N-(4-Pyridyl)-1,2-isothiazolidine-1,1-dioxide (4n)

To a solution of 3-chloropropylsulfonyl chloride 1 (3 ml, 24.7 mmol) and4-aminopyridine (2.32 g, 24.7 mmol) in DMF (25 ml) was slowly addedsodium hydride (60% in oil, 2.17 g, 54.3 mmol) over about 5 minutes withstirring under ice-cooling. The stirring was then continued for another30 minutes at 50° C. The reaction mixture was concentrated in vacuo, theresidue was subjected to column chromatography on silica gel. From thefraction eluted with a mixture of methylene chloride/methanol (10:1),1.294 (27%) of the desired compound 4n was obtained as a yellow solid.IR (ClCl₃)cm⁻¹: 3024, 2956, 1597, 1504, 1320, 1143. NMR (CDCl₃) δ:2.53-2.67 (2H, m, CH₂), 3.43 (2H, t, J=7.6 Hz, CH₂), 3.81 (2H, t, J=6.6Hz, CH₂), 7.08 (2H, d, J=5.4 Hz, CH), 8.49 (2H, d, J=5.4 Hz, CH).

Preparation 15 (R³=Et)(E)-2-Ethyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Ia)

To diisopropylamine (15.5 ml, 110.6 mmol) was dropwise added in anice-water bath n-butyllithium in n-hexane (1.6 M, 69.5 ml, 111 mmol)over 20 minutes with stirring. After completion of the addition,stirring was conducted for another 15 minutes. The reaction mixture wascooled to −78° C. and added THF (100 ml). To the reaction mixture wasdropwise added a solution of N-ethyl-1,2-isothiazolidine-1,1-dioxide 4a(15 g, 100.5 mmol), 3,5-di-tert-butyl-4-methoxymethoxybenzaldehyde 6a(25 g, 90.5 mmol) and HMPA (30 ml) in the THF (70 ml) over 15 minuteswith stirring. The reaction mixture was stirred for another 30 minutesat the same temperature, warmed to room temperature, poured into cold 2NHCl (100 ml) and extracted with ethyl acetate (2×250 ml). The ethylacetate phase was washed with a dilute aqueous solution of sodiumbicarbonate (300 ml) and a saturated brine (300 ml), dried overanhydrous sodium sulfate. The solvent was removed in vacuo, and theresidue was subjected to column chromatography on silica gel elutingwith n-hexane/ethyl acetate (4:1 to 1:1) to give 21.3 g (55%) of aldoladduct 7a as a colorless solid.

To a solution of the aldol adduct 7a (8.5 g, 19.9 mmol) in toluene (150ml) was added p-toluenesulfonic acid hydrate (2.49 g, 13 mmol). Theresultant mixture was heated to reflux for 30 minutes and then pouredinto a dilute aqueous solution of sodium bicarbonate (150 ml) andextracted with ethyl acetate (150ml×2). The organic layer was washedwith water (150 ml) and a saturated brine (150 ml), dried over anhydroussodium sulfate. The solvent was removed in vacuo, and the residue wassubjected to column chromatography on silica gel. From the fractioneluted with n-hexane/ethyl acetate (3:1), the desired compound Ia (2.59g, 36%) was yielded. M.p.=135-137° C. IR (KBr)cm⁻¹: 3610, 3440, 2970,2880, 1645, 1597, 1430, 1290, 1173, 1151, 1139. NMR (CDCl₃) δ: 1.29 (3H,t, J=7.2 Hz, CH₃), 1.45 (18H, s, 2×But), 3.07-3.19 (4H, m, CH₂), 3.28(2H, q, J=7.2 Hz, CH₂), 5.50 (1H, s, OH), 7.24-7.26 (3H, m,2×aromatic-H, CH). Elementary analysis (C₂₀H₃₁NO₃S) Calcd: C, 65.71; H,8.55; N, 3.83; S, 8.77. Found: C, 65.65; H, 8.43; N, 3.85; S, 8.78.

Preparation 16 (R³=CH₃)(E)-2-Methyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Ib)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (3.34 g, 12 mmol) andN-methyl-1,2-isothiazolidine-1,1-dioxide 4b (1.35 g, 10 mmol) to give1.65 g (40%) of adduct 7b. To a solution of the adduct 7b (1.60 g, 3.87mmol) in toluene (30 ml) was added p-toluenesulfonic acid hydrate (160mg) and the resultant mixture was heated to reflux for 30 minutes. Thereaction product was subjected to column chromatography on silica gel.From the fraction eluted with a mixture of n-hexane/ethyl acetate (3:7),the desired compound Ib (580 mg, 43%) was obtained. M.p.=168-170° C. IR(CHCl₃)cm⁻¹: 3620, 2956, 1435, 1292, 1218, 1149. NMR (CDCl₃) δ: 1.45(18H, s, 2×But), 2.76 (3H, s, NCH₃), 3.07-3.18 (2H, m, CH₂), 3.20-3.32(2H, m, CH₂), 5.51 (1H, s, OH), 7.23-7.29 (3H, m, 2×aromatic-H, CH).Elementary analysis (C₁₉H₂₉NO₃S) Calcd: C, 64.92; H, 8.32; N, 3.98; S,9.12. Found: C, 64.62; H, 8.31; N, 3.95; S, 9.14.

Preparation 17 (R³=CH₂CH(CH₃)₂)(E)-2-Isobutyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Ic)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (2.78 g, 10 mmol) andN-isobutyl-1,2-isothiazolidine-1,1-dioxide 4c (1.95 g, 11 mmol) to give3.67 g (81%) of adduct This adduct 7c (3.60 g, 7.9 mmol) was treated intoluene (50 ml) with p-toluenesulfonic acid hydrate (360 mg) in asimilar manner as that of Preparation 15. The product was subjected tocolumn chromatography on silica gel. From the fraction eluted with amixture of n-hexane-ethyl acetate (1:3), the desired compound Ic (1.30g, 42%) was obtained. M.p.=167-170° C. IR (CHCl₃)cm⁻¹: 3620, 2956, 1646,1435, 1289, 1240, 1148, 1081. NMR (CDCl₃) δ: 0.97 (6H, d, J=6.4 Hz,(CH₃)₂) 1.45 (18H, s, 2×But), 1.81-2.02 (1H, m, CH), 2.87 (2H, d, J=7.4Hz, CH₂), 3.06-3.18 (2H, m CH₂), 3.22-3.33 (2H, m, CH₂), 5.50 (1H, s,OH), 7.23-7.27 (3H, m, 2×aromatic-H, CH). Elementary analysis(C₂₂H₃₅NO₃S) Calcd: C, 67.14; H, 8.96; N, 3.56; S, 8.15. Found: C,66.85; H, 8.99; N, 3.58; S, 8.11.

Preparation 18 (R³=Cyclopropyl)(E)-2-Cyclopropyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Id)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (2.67 g, 9.6 mmol) andN-cyclopropyl-1,2-isothiazolidine-1,1-dioxide 4d (1.29 g, 8.0 mmol) togive 3.09 g (88%) of adduct 7d. The adduct 7d (3.0 g, 7 mmol) wastreated in toluene (50 ml) together with p-toluenesulfonic acid hydrate(300 mg). The reaction product was purified in a similar manner as thatof Preparation 17 to give 1.03 g (40%) of the desired compound Id.M.p.=202-204° C. IR (CHCl₃)cm⁻¹: 3620, 2956, 1434, 1297, 1237, 1145. NMR(CDCl₃) δ: 0.68-0.90 (4H, m, 2×CH₂), 1.44 (18H, s, 2×But), 2.28-2.40(1H, m, CH), 3.08 (2H, dt, J=2.6, 6.7 Hz, CH₂), 3.36 (2H, t, J=6.7 Hz,CH₂), 5.51 (1H, s, OH), 7.20-7.25 (3H, m, 2×aromatic-H, CH). Elementaryanalysis (C₂₁H₃₁NO₃S) Calcd: C, 66.81; H, 8.28; N, 3.71; S, 8.49. Found:C, 66.67; H, 8.29; N, 3.71; S, 8.38.

Preparation 19 (R³=CH₂CH₂CH₃)(E)-2-n-Propyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Ie)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (2.78 g, 10 mmol) andN-n-propyl-1,2-isothiazolidine-1,1-dioxide 4e (1.35 g, 8.27 mmol) togive 1.5 g (41%) of adduct 7e. The adduct 7e was treated withp-toluenesulfonic acid hydrate (400 mg) in a manner as described inPreparation 15. The reaction product was subjected to columnchromatography on silica gel. From the fraction eluted with a mixture ofn-hexane-ethyl acetate (1:4), the desired compound Ie (810 mg, 26%) wasobtained. M.p.=181-183° C. IR (CHCl₃)cm⁻¹: 3616, 2954, 1435, 1289, 1146.NMR (CDCl₃)cm⁻¹: 0.98 (3H, t, J=7.4 Hz, CH₃), 1.45 (18H, s, 2×But),1.57-1.78 (2H, m, CH₂), 2.98-3.20 (4H, m, 2×CH₂), 3.22-3.34 (2H, m,CH₂), 5.50 (₁H, s, OH), 7.23-7.27 (3H, m, 2×aromatic-H, CH ). Elementaryanalysis (C₂₁H₃₃NO₃S) Calcd: C, 66.45; H, 8.76; N, 3.69; S, 8.45. Found:C, 66.25; H, 8.74; N, 3.70; S, 8.33.

Preparation 20 (R³=OCH₃)(E)-2-Methoxy-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide (If) and Its (Z)-Isomer (9f)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (5.56 g, 20 mmol) andN-methoxy-1,2-isothiazolidine-1,1-dioxide 4f (3.32 g, 22 mmol) to give6.89 g (80%) of adduct 7f. The adduct 7f (6.89 g, 16 mmol) was treatedin toluene (100 ml) with p-toluenesulfonic acid hydrate (1 g) in amanner as described in Preparation 15. The reaction product wassubjected to column chromatography on silica gel. From the fractioneluted with a mixture of n-hexane-ethyl acetate (6:1), the desiredcompound If (2.40 g, 41%) was obtained. M.p.=166-168° C. IR (CHCl₃)cm⁻¹: 3616, 2952, 1639, 1436, 1340, 1240, 1158, 1002. NMR (CDCl₃) δ:1.45 (18H, s, 2×But), 3.11 (2H, dt, J=2.2, 7.0 Hz, CH₂), 3.66 (2H, t,J=7 Hz, CH₂), 3.81 (3H, s, OCH₃), 5.55 (1H, s, OH), 7.25-7.35 (3H, m,3×aromatic-H, CH). Elementary analysis (C₁₉H₂₉NO₄S) Calcd: C, 62.10; H,7.95; N, 3.81; S, 8.72. Found: C, 61.90; H, 7.88; N, 3.91; S, 8.67.

Preparation 21 (R³=OCH₂C₆H₅)(E)-2-Benzyloxy-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Ig)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (15 g, 54 mmol) andN-benzyloxy-1,2-isothiazolidine-1,1-dioxide 4g (10.23 g, 45 mmol) togive 15.51 g (68%) of adduct 7g. The adduct 7g (10.21 g, 20.2 mmol) wastreated in toluene (150 ml) with p-toluenesulfonic acid hydrate (1 g) ina manner as described in Preparation 15. The reaction product wasfiltered through a small amount of silica gel, and the filtrate wasconcentrated in vacuo to give 5.32 g (59%) of the desired compound Ig.M.p.=134-135° C. IR (CHCl₃)cm⁻¹: 3620, 2956, 1639, 1436, 1339, 1241,1159. NMR (CDCl₃) δ: 1.44 (18H, s, 2×But), 3.09 (2H, dt, J=2.6, 6.8 Hz,CH₂), 3.58 (2H, t, J=6.8 Hz, CH₂), 5.02 (2H, s, OCH₂), 5.53 (1H, s, OH),7.25-7.45 (8 H, m, 7×aromatic-H, CH). Elementary analysis (C₂₅H₃₃NO₄S)Calcd: C, 67.69; H, 7.50; N, 3.16; S, 7.23. Found: C, 67.52; H, 7.59; N,3.18; S, 7.16.

Preparation 22 (R³=4-Methoxybenzyl)(E)-2-(4-Methoxybenzyl)-5-(3,5-di-tert-butyl)benzylidene-1,2-isothiazolidine-1,1-dioxide(Ih)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (9 g, 32 mmol) andN-(4-methoxybenzyl)-1,2-isothiazolidine-1,1-dioxide Ah (7.24 g, 30 mmol)to give 13.61 g (84%) of adduct 7h. The adduct 7h (12.6 g, 24.2 mmol)was treated in toluene (150 ml) with p-toluenesulfonic acid hydrate (1.3g) in a manner as described in Preparation 15 to give 8.83 g of thedesired compound Ih.

Preparation 23 (R³=3,4-Dimethoxybenzyl)(E)-2-(3,4-Dimethoxybenzyl)-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Ii)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (5.6 g, 20 mmol) andN-(3,4-dimethoxy)benzyl-1,2-isothiazolidine-1,1-dioxide 4i (5.85 g, 21.6mmol) to give 9.25 g (78%) of adduct 7i. From the adduct 7i (4 g, 7.3mmol), the desired compound Ii (2.5 g) was obtained by dehydration anddeprotection in a manner as described in Preparation 15.

Preparation 24 (R³=C₆H₅)(E)-2-Phenyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Ij)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (2.47 g, 8.88 mmol) andN-phenyl-1,2-isothiazolidine-1,1-dioxide 4j (2.19 g, 11.10 mmol) to give3.184 g (75%) of adduct 7j. The adduct 7 J (3.184 g, 6.69 mmol) wastreated in toluene (100 ml) with p-toluenesulfonic acid hydrate (750 mg)to give the desired compound Ij (667 mg, 24%). M.p.=195-196° C. IR(KBr)cm⁻¹: 3560, 3520, 2960, 1636, 1593, 1492, 1430, 1295, 1268, 1105,1092. NMR (CDCl₃) δ: 1.47 (18H, s, 2×But), 3.31 (2H, dt, J=2.6, 6.6 Hz,CH₂), 3.80 (2H, t, J=6.6 Hz, CH₂), 5.54 (1H, s, OH), 7.17-7.26 (2H, m,aromatic-H, CH), 7.29 (2H, s, 2×aromatic-H), 7.38-7.42 (4H, m,4×aromatic-H). Elementary analysis (C₂₄H₃₁NO₃S×0.1 H₂O) Calcd: C, 69.39;H, 7.61; N, 3.37; S, 7.72. Found: C, 69.27; H, 7.60; N, 3.39; S, 7.61.

Preparation 25 (R³=4-Chlorophenyl)(E)-2-(4-Chlorophenyl)-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Ik)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (2.25 g, 8.09 mmol) andN-(4-chlorophenyl)-1,2-isothiazolidine-1,1-dioxide 4k (2.34 g, 10.1mmol) to give 2.54 g (62%) of adduct 7k. The adduct 7k (2.53 g, 4.96mmol) was treated in toluene (70 ml) with p-toluenesulfonic acid hydrate(250 mg) to give the desired compound Ik (859 mg, 39%). M.p.=245-246° C.IR (KBr)cm⁻¹: 3560, 2960, 1644, 1592, 1491, 1430, 1280, 1105, 1090. NMR(CDCl₃) δ: 1.46 (18H, s, 2×But), 3.30 (2H, dt, J=2.6, 6.6 Hz, CH₂), 3.76(2H, t, J=6.6 Hz, CH₂), 5.55 (1H, s, OH), 7.28 (2H, s, 2×aromatic-H),7.26-7.40 (5H, m, 4×aromatic-H, CH). Elementary analysis (C₂₄H₃₀NO₃SCl)Calcd: C, 64.34; H, 6.75; N, 3.13; S, 7.16; Cl, 7.91. Found: C, 64.59;H, 6.78; N, 3.28; S, 7.17; Cl, 7.87.

Preparation 26 (R³=2-Pyridyl)(E)-2-(2-Pyridyl)-5-(3,5-di-tert-butyl-4-hydroxyBenzylidene-1,2-isothiazolidine-1,1-dioxide (Il)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (208 mg, 0.75 mmol) andN-(2-pyridyl)-1,2-isothiazolidine-1,1-dioxide 4l (149 mg, 0.75 mmol) togive 233 mg (65%) of adduct 7l. The adduct 7l (231 mg, 0.485 mmol) wastreated in toluene (5 ml) with p-toluenesulfonic acid hydrate (60 mg) togive the desired compound Il (96 mg, 48%). M.p.=177-179° C. IR (KBr)cm¹:3570, 2960, 1646, 1600, 1587, 1472, 1431, 1300, 1105, 1085. NMR (CDCl₃)δ: 1.47 (18H, s, 2×But), 3.31 (2H, dt, J=2.4, 6.8 Hz, CH₂), 4.08 (2H, t,J=6.8 Hz, CH₂), 5.55 (1H, s, OH), 6.99-7.05 (1H, m, CH), 7.28 (2H, s,2×aromatic-H), 7.38 (1H, t, J=2.4 Hz, Py-H), 7.55-7.74 (2H, m, 2×Py-H),8.33-8.36 (1H, m, Py-H). Elementary analysis (C₂₃H₃₀N₂O₃S) Calcd: C,66.63; H, 7.29; N, 6.76; S, 7.73. Found: C, 66.31; H, 7.30; N, 6.72; S,7.66.

Preparation 27 (R³=3-Pyridyl)(E)-2-(3-Pyridyl)-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Im)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (1.474 g, 5.30 mmol) andN-(3-pyridyl)-1,2-isothiazolidine-1,1-dioxide 4m (1.051 g, 5.30 mmol) togive 1.522 g (60%) of adduct 7m. The adduct 7m (1.522 g, 3.19 mmol) wastreated in toluene (40 ml) with p-toluenesulfonic acid hydrate (400 mg)to give 358 mg (27%) of the desired compound Im. M.p.=207-209° C. IR(KBr)cm⁻¹: 3625, 3040, 2960, 1640, 1590, 1480, 1431, 1305, 1152. NMR(CDCl₃) δ: 1.47 (18H, s, 2×But), 3.36 (2H, dt, J=2.4, 6.4 Hz, CH₂), 3.84(2H, t, J=6.4 Hz, CH₂), 5.59 (1H, s, OH), 7.29 (2H, s, 2×aromatic-H),7.29-7.40 (2H, m, CH, Py-H), 7.84-7.93 (1H, m, Py-H), 8.37-8.64 (2H, m,2×Py-H). Elementary analysis (C₂₃H₃₀N₂O₃S) Calcd: C, 66.63; H, 7.29; N,6.76; S, 7.73. Found: C, 66.31; H, 7.27; N, 6.69; S, 7.47.

Preparation 28 (R³=4-Pyridyl)(E)-2-(4-Pyridyl)-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(In)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a (2.59 g, 9.36 mmol) andN-(4-pyridyl)-1,2-isothiazolidine-1,1-dioxide 4n (2.05 g, 10.4 mmol) togive 2.721 g (61%) of adduct 7n. The adduct 7n (1.65 g, 3.46 mmol) wastreated in toluene (80 ml) with p-toluenesulfonic acid hydrate (433 mg)to give 658 mg (46%) of the desired compound In. M.p.=213-214.5° C. IR(kBr)cm⁻¹: 3400, 2955, 1643, 1591, 1502, 1437, 1316, 1153. NMR (CDCl₃)δ: 1.47 (18H, s, 2×But), 3.37 (2H, dt, J=2.2, 6.8 Hz, CH₂), 3.82 (2H, t,J=6.8 Hz, CH₂), 5.61 (1H, s, OH), 7.21-7.25 (4H, m, 2×aromatic-H,2×Py-H), 7.42 (1H, t, J=2.2 Hz, CH), 8.50-8.58 (2H, m, 2×Py-H).Elementary analysis (C₂₃H₃₀N₂O₃S) Calcd: C, 66.63; H, 7.29; N, 6.76; S,7.73. Found: C, 66.46; H, 7.18; N; 6.66; S, 7.49.

Preparation 29 (R³=H) (Y=CO₂C (CH₃)₃)(E)-5-(3,5-Di-tert-Butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(10)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a andN-(tert-butoxycarbonyl)-1,2-isothiazolidine-1,1-dioxide 4o, which hadbeen prepared from starting materials, 3-chloropropylsulfonyl chlorideand tert-butyl carbonate, in accordance with the method of reactionscheme I, to give a crude adduct 7o. To a solution of the crude adduct7o in toluene was added p-toluenesulfonic acid hydrate and the resultantmixture was heated to reflux for 45 minutes and then subjected to columnchromatography on silica gel. From the fraction eluted with a mixture ofn-hexane/ethyl acetate (2:1), the desired compound 10 (yield 8.5%) wasobtained. M.p.=233-234° C. IR (CHCl₃)cm⁻¹: 3618, 2952, 1435, 1366, 1311,1240, 1155, 1070. NMR (CDCl₃) δ: 1.45 (18H, s, 2×But), 3.18 (2H, dt,J=2.6, 6.8 Hz, CH₂), 3.42-3.60 (2H, m, CH₂), 4.05-4.25 (1H, broad, NH),5.52 (1H, s, OH), 7.22-7.27 (3H, m, 2×aromatic-H, CH). Elementaryanalysis (C₁₈H₂₇NO₃S×0.35H₂O) Calcd: C, 62.89; H, 8.12; N, 4.07; S,9.38. Found: C, 63.10; H, 7.90; N, 4.17; S, 9.11.

Preparation 30 (R³=H) (Y=4-Methoxybenzyl)(Z)-5-(3,5-Di-tert-Butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(10)

To a solution of the adduct 7h (13.16 g, 25.3 mmol) of the aldolreaction, which was obtained in a manner as described in Preparation 22,in toluene (150 ml) was added p-toluenesulfonic acid hydrate (1.3 g).The resultant mixture was heated to reflux for 30 minutes and filteredthrough a small amount of silica gel. The solvent was removed in vacuato give a mixture (8.83 g) of crude (E)- and(Z)-2-(4-methoxybenzyl)-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxides.To a solution of the mixture in methylene chloride (150 ml) was addedtitanium tetrachloride (4.1 ml). The resultant mixture was stirred for30 minutes at 0° C. and then subjected to column chromatography onsilica gel. From the fraction eluted with a mixture of n-hexane/ethylacetate (1:1), compounds 10 (3.35 g, 41%) was obtained. Compound 10 wasidentical with the sample obtained in Preparation 29.

Preparation 31 (R³=H) (Y=3,4-Dimethoxybenzyl)

To a solution of the adduct 7i (4.0 g, 7.3 mmol) of the aldol reactionobtained in Preparation 9 in xylene (50 ml) were added an equimolaramount of each of 2,6-di-tert-butylphenol, anisole and p-toluenesulfonicacid hydrate. The resultant mixture was heated to reflux for 45 minutes,and the reaction product was subjected to column chromatography onsilica gel to give compounds 10 (580 mg, 24%). Compound 10 was identicalwith the samples obtained in Preparations 29 and 30, respectively.

Preparation 32 (R³=CH₂CO₂C₂H₅)(E)-2-Ethoxycarbonylmethyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Ip)

(E)-5-(3,5-Di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide10 (500 mg, 1.48 mmol), ethyl iodoacetate (240 pl, 2 mmol), an aqueoussolution of 2N sodium hydroxide (1.5 ml, 3 mmol) and a small amount ofN-benzyltrimethylammonium chloride were sequentially added to a mixtureof chloroform (20 ml) and water (10 ml). The resultant mixture wasstirred for 24 hours at room temperature and then treated in aconventional manner. The product was purified by column chromatographyon silica gel to give 300 mg (49%) of the desired compound Ip. IR(CHCl₃)cm⁻¹: 3620, 2956, 1747, 1435, 1298, 1229, 1160. NMR (CDCl₃) δ:1.29 (3H, t, J=7.2 Hz, CH₃), 1.45 (18H, s, 2×But), 3.19 (2H, dt, J=2.6,6.6 Hz, CH₂), 3.51 (2H, t, J=6.6 Hz, CH₂), 3.87 (2H, s, CH₂CO), 4.23(2H, q, J=7.2 Hz, CH₂), 5.52 (1H, s, OH), 7.22-7.30 (3H, m,2×aromatic-H, CH).

Preparation 33 (R³=CH₂COOH)(E)-2-Carboxymethyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Iq)

Compound Iq (610 mg, 1.44 mmol) as obtained in a similar manner asdescribed in Preparation 32 and an aqueous solution of 2N sodiumhydroxide (1.5 ml) were added to a mixture of THF (10 ml) and methanol(4 ml). The resultant mixture was stirred at 0° C. for 30 minutes. Afterthe addition of ethyl acetate (50 ml), the reaction mixture was washedwith an aqueous solution of 1N hydrochloric acid (20 ml), then asaturated brine (20 ml), and dried over anhydrous sodium sulfate. Thesolvent was removed in vacuo to give 445 mg (78%) of the desiredcompound Iq. M.p=175-178° C. IR (CHCl₃)cm⁻¹: 3620, 2954, 1735, 1435,1297, 1240, 1149. NMR (CDCl₃) δ: 1.45 (18H, s, 2×But), 3.20 (2H, dt,J=2.6, 6.6 Hz, CH₂), 3.51 (2H, t, J=6.6 Hz, CH₂), 3.95 (2H, s, CH₂CO),5.54 (1H, s, OH), 7.25 (2H, s, 2×aromatic-H). Elementary analysis(C₂₀H₂₉NO₅S) Calcd: C, 60.46; H, 7.41; N, 3.53; S, 8.07. Found: C,60.34; H, 7.40; N, 3.56; S, 8.04.

Preparation 34 (R³=CH₂CH₂OH)(E)-2-(2-Hydroxyethyl)-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1.1-dioxide(Ir)

Compound 10 (675 mg, 2 mmol), 2-iodoethanol (624 μl, 8 mmol), an aqueoussolution of 2N sodium hydroxide (2 ml) and a small amount ofN-benzyltrimethylammonium chloride were added to a mixture of methylenechloride (20 ml) and water (10 ml). The resultant mixture was heated toreflux for 3 days and treated in a conventional manner and the productwas subjected to column chromatography on silica gel. From the fractioneluted with a mixture of n-hexane/ethyl acetate (7:3), 190 mg (25%) ofthe desired compound Ir was obtained. M.p.=156-157° C. IR (CHCl₃)cm⁻¹:3620, 2950, 1434, 1290, 1240, 1151, 1066. NMR (CDCl₃) δ: 1.45 (18H, s,2×But), 3.16 (2H, dt, J=2.4, 6.5 Hz, CH₂), 3.30 (2H, m, CH₂), 3.41 (2H,t, J=6.5 Hz, CH₂), 3.87 (2H, t, J=5.2 Hz), 5.53 (1H, s, OH), 7.23-7.29(3H, m, 2×aromatic-H, CH). Elementary analysis (C₂₀H₃₁NO₄S) Calcd: C,62.96; H, 8.19; N, 3.67; S, 8.40. Found: C, 62.72; H, 8.27; N, 3.69; S,8.21.

Preparation 35 (R³=CH₂CH₂N(CH₃)₂)(E)-2-(2-Dimethylamino)ethyl)-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Is)

Compound 10 (843 mg, 2.5 mmol), N,N-dimethyl-2-bromoethylamine (750 mg,5 mmol), an aqueous solution of 2N sodium hydroxide (3 ml, 6 mmol) and asmall amount of N-benzyltrimethylammonium chloride were added to amixture of chloroform (30 ml) and water (10 ml). The resultant mixturewas stirred for 2 hours under ice-cooling. The chloroform layer waswashed with water (20 ml×2) and dried over anhydrous sodium sulfate.Chloroform was removed in vacuo to give 950 mg (93%) of the desiredcompound as a crystalline residue. M.p.=160-165° C. IR (CHCl₃)cm⁻¹:3620, 2956, 1435, 1290, 1148. NMR (CDCl₃) δ: 1.45 (18H, s, 2×But), 2.29(6H, s, N(CH₃)₂), 2.60 (2H, t, J=6.6 Hz, CH₂), 3.12 (2H, dt, J=2.2, 6.6Hz, CH₂), 3.20 (2H, t, J=6.6 Hz, CH₂), 3.38 (2H, t, J=6.6 Hz, CH₂), 5.51(1H, s, OH), 7.21-7.28 (3H, m, 2×aromatic-H, CH). Elementary analysis(C₂₂H₃₆N₂O₃S×0.2CH₂Cl₂) Calcd: C, 62.65; H, 8.62; N, 6.58; S, 7.53; Cl,3.33. Found: C, 62.32; H, 8.60; N, 6.71; S, 7.56; Cl, 3.24.

Preparation 36 (R³=COCH₃)(E)-2-Acetyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(It)

To a solution of compound 10 (585 mg, 1.74 mmol) in pyridine (10 ml), asmall amount of 4-N,N-dimethylaminopyridine was added, and aceticanhydride (6 ml) was dropwise added under ice-cooling. The resultantmixture was stirred for 1 hour at room temperature. The mixture wasconcentrated under reduced pressure, and the residue was dissolved inethyl acetate and the solution was filtered through a small amount ofsilica gel. The solvent was removed in vacuo to give 360 mg (55%) of thedesired compound as a crystal-like residue. M.p.=177-179° C. IR(CHCl₃)cm⁻¹: 3618, 2958, 1695, 1435, 1379, 1297, 1153, 1117. NMR (CDCl₃)δ: 1.46 (18H, s, 2×But), 2.53 (3H, s, COCH₃), 3.20 (2H, dt, J=2.2, 7.0Hz, CH₂), 3.86 (2H, t, J=7.0 Hz, CH₂), 5.60 (1H, s, OH), 7.52 (2H, s,2×aromatic-H), 7.39 (1H, t, J=2.2 Hz, CH). Elementary analysis(C₂₀H₂₉NO₄S) Calcd: C, 63.30; H, 7.70; N, 3.69; S, 8.45. Found: C,63.27; H, 7.83; N, 3.64; S, 8.22.

Preparation 37 ((R³=N-methyl-N-methoxy)carbamoyl)(E)-2-(N-Methyl-N-methoxy)carbamoyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Iu)

Compound 10 (450 mg, 1.33 mmol) and N-methyl-N-methoxy-O-phenylcarbamate (300 mg, 1.66 mmol) were dissolved in a mixture of THF (10 ml)and HMPA (10 ml). To the solution was dropwise added a solution oflithium hexamethyldisilazane (LiHMDS) in THF (1 M, 3.2 ml) with stirringand cooling to −40° C. The reaction mixture was warmed to roomtemperature and poured into an aqueous solution of 1N hydrochloric acid(20 ml). The mixture was extracted with ethyl acetate (30 ml), and thenthe ethyl acetate layer was washed with water (30 ml) and a saturatedbrine (30 ml), dried over anhydrous sodium sulfate. The solvent wasremoved under reduced pressure, and the residue was subjected to columnchromatography on silica gel. From the fraction eluted with a mixture ofn-hexane/ethyl acetate (7:3), the desired compound Iu (230 mg, 41%) wasobtained. IR (CHCl₃)cm⁻¹: 3620, 2958, 1673, 1435, 1388, 1330, 1240,1207, 1155, 1092. NMR (CDCl₃) δ: 1.45 (18H, s, 2×But), 3.21 (2H, dt,J=2.2, 6.8 Hz, CH₂), 3.31 (3H, s, NCH₃), 3.78 (3H, s, OCH₃), 3.89 (2H,t, J=6.8 Hz), 5.54 (1H, s, OH), 7.23 (2H, s, 2×aromatic-H), 7.31 (1H, t,J=2.2 Hz, CH).

Preparation 38 ((R³=N-Benzyloxy-N-methoxymethyl)carbamoyl)(E)-2-(N-Benzyloxyl-N-methoxymethyl)carbamoyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Iv)

Compound 10 (424 mg, 1.26 mmol) and N-benzyloxy-N-methoxymethyl-O-phenylcarbamate (722 mg, 2.52 mmol) were treated in a mixture of THF (90 ml)and HMPA (30 ml) with a solution of LiHMDS in THF (1 M, 4.0 ml), in amanner as described in Preparation 37. The reaction product wassubjected to column chromatography on silica gel. From the fractioneluted with a mixture of n-hexane/ethyl acetate (3:1), the desiredcompound Iv (600 mg, 90%) was obtained.

NMR (CDCl₃) δ: 1.45 (18H, s, 2×But), 3.18 (2H, dt, J=2.0, 6.8 Hz, CH₂),3.45 (3H, s, OCH₃), 3.79 (2H, t, J=6.8 Hz, CH₂), 4.94 (2H, s, OCH₂),5.02 (2H, s, OCH₂), 5.54 (1H, s, OH), 7.22 (2H, s, 2 x aromatic-H), 7.30(1H, t, J=2.0 Hz, CH), 7.30-7.55 (5H, m, 5×aromatic-H).

Preparation 39 (R³=CONHOH)(E)-2-(Hydroxycarbamoyl)-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1.2-isothiazolidine-1,1-dioxide(Iw)

To a solution of compound Iv (600 mg, 1.13 mmol) obtained in Preparation38 in methylene chloride (8 ml) was added titanium tetrachloride (500μl, 4.56 mmol) under ice-cooling, and the resultant mixture was stirredfor 1.5 hours. After an aqueous solution of 2N hydrochloric acid (10 ml)was added, the reaction mixture was stirred for 30 minutes at roomtemperature and then extracted with methylene chloride (20 ml). Theorganic layer was washed with a saturated brine (20 ml), dried overanhydrous sodium sulfate. The solvent was removed under reducedpressure, and the residue was subjected to column chromatography onsilica gel. From the fraction eluted with a mixture of n-hexane/ethylacetate (1:1), the desired compound Iw (150 mg, 33%) was obtained. IR(3.94 (2H, t, J=7.0 Hz, CH₂), 5.61 (1H, s, OH), 6.85-6.95 (1H, broad,OH), 7.24 (2H, s, 2×aromatic-H), 7.30 (1H, t, J=2.2 Hz, CH), 8.61 (1H,s, NH).

Preparation 40(E)-2-Hydroxy-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(12)

According to a similar method to that of Preparation 15, an aldolreaction was carried out using compound 6a andN-benzyloxy-1,2-isothiazblidine-1,1-dioxide. The adduct obtained by thealdol reaction was then treated with p-toluenesulfonic acid hydrate togive crude2-benzyloxy-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide11. To a solution of the crude dioxide (4.44 g, 10 mmol) in methylenechloride (80 ml) was dropwise added titanium tetrachloride (4.4 ml, 40mmol) with stirring and ice-cooling, and the mixture was stirred foranother 2 hours at the same temperature. An aqueous solution of 1Nhydrochloric acid (50 ml) was added to the reaction mixture. Themethylene chloride layer was separated, washed sequentially with water(50 ml) and a saturated brine (50 ml), dried over anhydrous sodiumsulfate. The solvent was removed under reduced pressure, and the residuewas subjected to column chromatography on silica gel. From the fractioneluted with a mixture of n-hexane/ethyl acetate (3:1), the desiredcompound 12 (1.6 g, 45%) was obtained. M.p.=177-182° C. (decomp.). IR(KBr)cm⁻¹: 3560, 3430, 1425, 1330, 1240, 1155, 1130, 1115. NMR (CDCl₃)δ: 1.45 (18H, s, 2×But), 3.18 (2H, dt, J=2.6, 6.8 Hz, CH₂), 3.89 (2H, t,J=6.8 Hz, CH₂), 5.56 (1H, s, OH), 6.18-6.30 (1H, broad, OH), 7.26-7.35(3H, m, 2×aromatic-H, CH). Elementary analysis (C₁₈H₂₇NO₄S) Calcd: C,61.16; H, 7.70; N, 3.96; S, 9.07. Found: C, 60.86; H, 7.68; N, 3.93; S,8.90.

Preparation 41 (E)-2-Isopropyl-5-(3,5-di-tert-butyl-4-hydroxy)benzylidene-1,2-isothiazolidine-1,1-dioxide(Ix)

According to a similar method to that of Preparation 15, an aldolreaction was carried out usingN-isopropyl-1,2-isothiazolidine-1,1-dioxide 4p (3.65 g, 22.4 mmol) and3,5-di-tert-butyl-4-methoxymethoxybenzaldehyde 6a (5.28 g, 19.0 mmol) togive 6.27 g (74.7%) of adduct 7p as a white powder. To a solution of theadduct 7p (6.27 g) in toluene (120 ml) was added p-toluenesulfonic acidhydrate (600 mg). The mixture was heated to reflux for 30 minutes,cooled, washed twice with water (100 ml), dried over anhydrous sodiumsulfate, and the solvent was removed under reduced pressure. Thecrystalline residue was recrystallized from methanol to give 2.16 g(30%) of the desired compound Ix as colorless columnar crystals.M.p.=148-150° C. IR (KBr)cm⁻¹: 3550, 2960, 1645, 1600, 1432, 1273, 1173.NMR (CDCl₃) δ: 1.29 (6H, d, J=6.6 Hz, 2×CH₃), 1.45 (18H, s, 2×But),3.07-3.14 (2H, m, CH₂), 3.29-3.35 (2H, m, CH₂), 3.94 (1H, sept, CH),5.48 (1H, s, OH), 7.22 (1H, t, J=2.8 Hz, CH), 7.23 (2H, s, Ar—H).Elementary analysis (C₂₁H₃₃NO₃S) Calcd: C, 66.45; H, 8.76; N, 3.69; S,8.45. Found: C, 66.37; H, 9.01; N, 3.67; S, 8.28.

Preparation 42 (R³=CH₂CH₂CH₃) N-n-Propyl-1,2-isothiazolidine-1,1-dioxide(4e)

To γ-sultone (12.2 g, 0.1 mol) was added n-propylamine (5.9 g, 0.1 mmol)with stirring and ice-cooling. As the reaction proceeds, the contentssolidified. To the solid product was added phosphorus oxychloride (10ml). The reaction mixture was heated to reflux for 2 hours, and theremaining phosphorus oxychloride was removed under reduced pressure.After the addition of ether (100 ml) to the residue, insolublesubstances were removed by filtration. The ether layer was dried overanhydrous sodium sulfate, and the solvent was removed in vacuo to givethe desired compound 4e (15.2 g, 93%) as a colorless oil. This productwas identical with the sample obtained in Preparation 5.

Preparation 43 (R³=CH₂CH(CH₃)₂)N-Isobutyl-1,2-isothiazolidine-1,1-dioxide (4c)

According to a similar method to that of Preparation 1, γ-sultone (12.2g, 0.1 mol), isobutylamine (7.3 g, 0.1 mol) and phosphorus oxychloride(10 ml) were reacted to give the desired compound 4c (15.9 g, 90%). Thisproduct was identical with the sample obtained in Preparation 3.

Novel compounds of the present invention can be prepared according tothe following Examples, and the processes for the production describedtherein per se are not to be construed as limiting the presentinvention. One example of the processes for the production of the novelcompounds of the present invention is provided below.

EXAMPLES Example 1

a. 3,5-Diallyl-4-hydroxybenzaldehyde (23)

The titled compound was synthesized according to the process describedin the literature with a partial modification. See Claisen; Eisleb,Justus Liebigs Ann. Chem., 401, 108 (1913).

b.2,6-Diallyl-4-((2-ethyl-1,1-dioxo-1,2-isothiazolidin-5-yl)hydroxy-methyl)phenol(24)

LDA (2.0 M solution in THF) (4.52 ml, 9.04 mmol) was dropwise and slowlyadded to a solution of 23 (850 mg, 4.20 mmol) and2-ethyl-1,2-isothiazolidin-1,1-dioxide (752 mg, 5.04 mmol) in THF (15ml), while maintaining the temperature below −45° C. The reactionmixture was warmed to 0° C. and stirred for another 1 hour. The mixturewas added with diluted hydrochloric acid and extracted with ethylacetate. The organic layer was washed with water and saturated brine,dried over anhydrous sodium sulfate, and the solvent was removed invacuo. The residue was purified by column chromatography on silica gelto yield 1.083 g (73%) of 24 as a mixture of diastereomers: ¹H NMR(CDCl₃) δ: 1.23-1.28 (3H, m), 1.75-2.06 (2H*1/2, m), 2.47-2.61 (2H*1/2,m), 2.95-3.53 (9H, m), 4.79-5.42 (6H, m), 5.90-6.08 (2H, m), 7.02 (2H,s).

c.5-((tert-Butyl-dimethylsilanyloxy)-(3,3-diallyl-4-methoxymethoxyphenyl)-methyl)-2-ethyl-1,2-isothazolidin1,1-dioxide (25)

To a solution of 24 (1.789 g, 5.09 mmol) in dichloromethane (25 ml),under ice-cooling, diisopropylethylamine (4.43 ml, 25.45 mmol) and MOMCl(1.16 ml, 15.27 mmol) were dropwise added sequentially and slowly. Afterstirring at 0° C. for 1 hour and then at room temperature for 1 hour,the reaction mixture was added with diluted hydrochloric acid andextracted with chloroform. The organic layer was washed with water,saturated sodium bicarbonate and saturated brine. After drying overanhydrous sodium sulfate, the solvent was removed in vacuo. The residuewas purified by column chromatography on silica gel to yield 1.470 g(73%) of an intermediate. This intermediate (805 mg, 2.035 mmol) wasdissolved in dichloromethane (10 ml), 2,6-lutidine (0.71 ml, 6.105 mmol)and TBSOTf (0.70 ml, 3.05 mmol) were added to the solution underice-cooling, and the mixture was stirred for 2 hours. The mixture wasadded with diluted hydrochloric acid and extracted with chloroform. Theorganic layer was washed with water, saturated sodium bicarbonate andsaturated brine. After drying over anhydrous sodium sulfate, the solventwas removed in vacuo. The residue was purified by column chromatographyon silica gel to yield 960 mg (92%) of 25. ¹H NMR (CDCl₃) δ: −0.25(3H*1/2, s), −0.23 (3H*1/2, s), 0.08 (3H*1/2, s), 0.11 (3H*1/2, s), 0.83(9H*1/2, s), 0.86 (9H*1/2, s), 1.20 (3H, t, J=7.2 Hz), 1.60-2.54 (2H,m), 2.91-3.52 (9H, m), 3.58 (3H*1/2, s), 3.59 (3H*1/2, s), 4.80-5.12(7H, m), 5.85-6.04 (2H, m), 7.03 (2H*1/2, s), 7.06 (2H*1/2, s).

d. Methyl(5-((2-Ethyl-1,1-dioxo-1,2-isothiazolidin-5-yl)-(tert-butyl-dimethylsilanyloxy)-methyl)-3-methoxycarbonylmethyl-2-methoxymethoxy-phenyl)acetate(26)

Ozone gas was passed through a solution of 25 (3.90 g, 7.65 mmol) indichloromethane (100 ml) and methanol (20 ml) at −780° C. until thecolor of the solution turned to blue. To the reaction mixture,triethylphosphite (3.94 ml, 23 mmol) was added, and the mixture wasslowly warmed to room temperature. The reaction mixture wasconcentrated, and the residue was added with t-butanol (40 ml), H₂O (10ml), 2-methyl-2-butene (8.1 ml, 76.5 mmol) and NaH₂PO₄ (2.31 g, 15.3mmol). NaClO₂ (3.50 g, 30.6 mmol) was then slowly added to the mixtureat room temperature. After stirring overnight, the mixture was addedwith hydrochloric acid and extracted with chloroform. The organic layerwas washed with saturated brine, dried and concentrated. The resultantresidue was added with ethyl acetate, and excess amount of an ethersolution of diazomethane was dropwise added to the mixture underice-cooling. The reaction mixture was concentrated and purified bycolumn chromatography on silica gel to yield 2.187 g (50%) of 26. ¹H NMR(CDCl₃) δ: −0.23 (3H*1/2, s), −0.21 (3H*1/2, s), 0.09 (3H*1/2, s), 0.12(3H*1/2, s), 0.84 (9H*1/2, s), 0.86 (9H*1/2, s), 1.19 (3H*1/2, t, J=7.2Hz), 1.34 (3H*1/2, t, J=7.2 Hz), 1.62-2.58 (2H, m), 2.90-3.96 (14H, m),4.85-5.13 (7H, m), 7.16 (2H*1/2, s), 7.21 (2H*1/2, s).

e. Methyl2-(5-((2-Ethyl-1,1-dioxo-1,2-isothiazolidin-5-yl)-(tert-butyl-dimethylsilanyloxy)-methyl)-3-(1-methoxycarbonyl-1-methyl-ethyl)-2-methoxymethoxy-phenyl)-2-methyl-propionate(27a and 27b)

A solution of 26 (4.54 g, 7.91 mmol) and MeI (3.95 ml, 63.3 mmol) in DMF(25 ml) was slowly and dropwise added to a suspension of NaH (60% inmineral oil, 1.26 g, 31.6 mmol) (n-hexane wash) in DMF (5 ml) underice-cooling. After stirring overnight at room temperature, the reactionmixture was added with diluted hydrochloric acid and extracted withethyl acetate. The organic layer was washed with water and saturatedbrine. After drying over anhydrous sodium sulfate, the solvent wasremoved to obtain a residue (3.90 g). To this residue, THF (40 ml) wasadded, and LiHMDS (1.0 M in THF) (14.25 ml, 14.25 mmol) was dropwiseadded over 10 minutes under ice-cooling. After stirring for another 45minutes, the mixture was added with MeI (3.24 ml, 51.8 mmol) and warmedto room temperature. After stirring for 1.5 hours at room temperature,the mixture was added with diluted hydrochloric acid and extracted withethyl acetate. The organic layer was washed with water and saturatedbrine. After drying over anhydrous sodium sulfate, the solvent wasremoved in vacuo. The residue was purified by column chromatography onsilica gel to yield 2.50 g (48%) of the titled compound as a mixture ofdiastereomers ((27a) less polar, 1.08 g, (27b) more polar 1.42 g). ¹HNMR (CDCl₃) δ (27a):−0.20 (3H, s), 0.11 (3H, s), 0.88 (9H, s), 1.22 (3H,t, J=7.2 Hz), 1.53 (6H, s), 1.55 (6H, s), 2.44-2.60 (2H, m), 2.92-3.40(5H, m), 3.49 (3H, s), 3.64 (6H, s), 4.78 (2H, ABq, J=4.5, 6.6 Hz), 5.18(1H, d, J=4.8 Hz), 7.28 (2H, s). ¹H NMR (CDCl₃) δ (27b):−0.23 (3H, s),0.12 (3H, s), 0.82 (9H, s), 1.21 (3H, t, J=7.2 Hz), 1.53 (6H, s), 1.58(6H, s), 1.70-1.89 (2H, m), 2.94-3.67 (14H, m), 4.77 (2H, s), 4.80-4.92(1H, m), 7.23 (2H, s).

f. Methyl2-(5-((2-Ethyl-1,1-dioxo-1,2-isothiazolidin-5-yl)-hydroxy-methyl)-3,3-dimethyl-2-oxo-2,3-dihydro-benzofuran-7-yl)-2-methyl-propionate(28)

To a solution of 27a (416 mg, 066 mmol) in methanol (6 ml), 4NHCl/dioxane (6 ml) was added under ice-cooling, and the mixture wasstirred for 2 hours at room temperature. After the reaction mixture wasconcentrated, the mixture was added with water and extracted with ethylacetate. The organic layer was washed with water and saturated brine,dried, and concentrated. The residue was purified by columnchromatography on silica gel to yield 207 mg (71%) of 28. ¹H NMR (CDCl₃)δ: 1.27 (3H, t, J=7.2 Hz), 1.49 (6H, s), 1.61 (6H, s), 1.97-2.10 (1H,m), 2.46-2.61 (1H, m), 3.02-3.80 (8H, m), 5.49 (1H, brs), 7.17 (1H, s),7.20 (1H, s).

g. Methyl2-(5-(2-Ethyl-1,1-dioxo-1,2-isothazolidin-5-ylidenemethyl)-3,3-dimethyl-2-oxo-2,3-dihydro-benzofuran-7-yl)-2-methyl-propionate(29)

To a solution of 28 (318 mg, 0.724 mmol) and Ph₃P (569 mg, 2.17 mmol) inTHF (5 ml), DEAD (0.171 ml, 1.09 mmol) was slowly added underice-cooling. After stirring for 1 hour, the reaction mixture wasconcentrated, and the residue was purified by column chromatography onsilica gel to yield 183 mg (60%) of 29. Mp.116-120° C. (colorlesspowder). ¹H NMR (CDCl₃) δ: 1.30 (3H, t, J=7.2 Hz), 1.51 (6H, s), 1.63(6H, s), 3.10-3.22 (4H, m), 3.31 (2H, t, J=6.3 Hz), 7.15 (1H, d, J=1.8Hz), 7.26-7.29 (2H, m).

h.2-(5-(2-Ethyl-1,1-dioxo-1,2-isothiazolidin-5-ylidenemethyl)-2-hydroxy-3,3-dimethyl-2,3-dihydro-benzofuran-7-yl)-2-methyl-propionicacid (30)

2N KOH (1.5 ml) was added to a solution of 29 (19 mg, 0.0364 mmol) inmethanol (2 ml), and the mixture was allowed to stand overnight at roomtemperature. The reaction mixture was concentrated, acidified withdiluted hydrochloric acid, and extracted with ethyl acetate. The organiclayer was washed with saturated brine, and after dryness, the solventwas removed in vacuo. The residue was added with dichloromethane (1 ml),cooled to −78° C., and slowly and dropwise added with DIBAL (0.93 M inn-hexane) (0.31 ml, 0.182 mmol). After 5 minutes, a saturated aqueousammonium chloride solution was added to quench the reaction, dilutedhydrochloric acid was added, and the mixture was extracted with ethylacetate. The organic layer was washed with saturated brine, dried, andconcentrated. The residue was purified by preparative TLC to yield 5.9mg (32%) of 30. ¹H NMR (d6-aceton) δ: 1.23 (3H, t, J=7.3 Hz), 1.29 (6H,br), 1.57 (6H, s), 3.06 (2H, q, J=7.3 Hz), 3.19 (2H, td, J=6.5, 2.6 Hz),3.31 (2H, t, J=6.5 Hz), 5.66 (1H, s), 6.71 (1H, br), 7.12 (1H, t, J=2.6Hz), 7.27 (1H, d, J=1.6 Hz), 7.34 (1H, d, J=1.6 Hz).

i.5-(2-Ethyl-1,1-dioxo-1,2-isothiazolidin-5-ylidenemethyl)-7-(2-hydroxy-1,1-dimethyl-ethyl)-3,3-dimethyl-2,3-dihydro-benzofuran-2-ol(31)

To a solution of 29 (56 mg, 0.133 mmol) in hexane (1 ml) anddichloromethane (1 ml) at −78° C., DIBAL (0.93 M) (0.64 ml, 0.595 mmol)was slowly added. The mixture was stirred for another 1 hour under thesame conditions. The reaction mixture was added with dilutedhydrochloric acid, warmed to room temperature, and stirred for another15 minutes at room temperature. The resultant product was extracted withethyl acetate, washed with water, saturated sodium bicarbonate, andsaturated brine, dried, and concentrated. The residue was purified bycolumn chromatography over silica gel to yield 31 mg (59%) of 31 asamorphous solid. Mp.78-82° C. ¹H NMR (d6-aceton) δ: 1.23 (3H, t, J=7.3Hz), 1.26 (3H, brs), 1.34 (3H, brs), 1.36 (6H, s), 3.05 (2H, q, J=7.3Hz), 3.19 (2H, m), 3.31 (2H, t, J=6.4 Hz), 3.59 (2H, t, J=5.6 Hz), 3.76(2H, m), 5.66 (1H, s), 6.27 (1H, br), 7.11 (1H t, J=2.7 Hz), 7.26 (1H,d, J=1.8 Hz), 7.36 (1H, d, J=1.8 Hz).

Example 2

a. Benzoic-acid2-(3-(2-Benzoyloxy-1,1-dimethyl-ethyl)-5-((tert-butyl-dimethyl-silanyloxy)-(2-ethyl-1,1-dioxo-1,2-isothiazolidin-5-yl)-methyl)-2-methoxymethoxy-phenyl)-2-methyl-propylester (32)

To a solution of 27b (529 mg, 0.84 mmol) in THF (8 ml), LiBH₄ (146 mg,6.72 mmol) was added, and the mixture was refluxed for 5 hours. Thereaction mixture was added with diluted hydrochloric acid and extractedwith ethyl acetate. The organic layer was dried and purified by columnchromatography on silica gel to yield 309 mg of diol. This diol (141 mg,0.246 mmol) was benzoylated according to a conventional method to give145 mg (39% for 2 steps) of 32. ¹H NMR (CDCl₃) δ: −0.27 (3H, s), 0.10(3H, s), 0.82 (9H, s), 1.46 (3H, t, J=7.2 Hz), 1.49 (12H, s), 2.60-3.32(7H, m), 3.64 (3H, s), 4.62 (4H, ABq, J=11, 25 Hz), 4.81 (1H, d, J=9.3Hz), 4.95 (2H, s), 7.26-7.87 (12H, m).

b.4-(2-Ethyl-1,1-dioxo-1,2-isothiazolidin-5-ylidenemethyl)-2,6-bis-(2-benzoyloxy-1,1,dimethyl-ethyl)-phenol(33)

To a solution of 32 (45 mg, 0.0575 mmol) in methanol (1 ml), 4NHCl/dioxane (1 ml) was added under ice-cooling, and the mixture waswarmed to room temperature and stirred for 2 hours. The mixture wastreated in the manner as described above, and benzene (1 ml) andcatalytic amount of p-TsOH were added to the residue, and the mixturewas refluxed for 3 hours. The reaction mixture was added with water andextracted with ethyl acetate. The organic layer was washed withsaturated brine, dried, and concentrated. The residue was purified bypreparative TLC to yield 12 mg (34%)of 33. ¹H NMR (CDCl₃) δ: 1.27 (3H,t, J=7.5 Hz), 1.54 (12H, s), 3.02 (2H, td, J=7.2. 2.7 Hz), 3.14 (2H, q,J=7.5 Hz), 3.18 (2H, t, J=7.2 Hz), 4.60 (4H, s), 7.24 (1H, t, J=2.7 Hz),7.31 (2H, s), 7.38-7.93 (10H, m).

c.4-(2-Ethyl-1,1-dioxo-1,2-isothiazolidin-5-ylidenemethyl)-2,6-bis-(2-hydroxy-1,1-dimethyl-ethyl)-phenol(34)

A solution of 33 (12 mg, 0.0198 mmol) in dichloromethane (1 ml) wascooled to −78° C., and DIBAL (0.93 M) (0.107 ml, 0.099 mmol) was slowlyadded. After 5 minutes, the mixture was added with diluted hydrochloricacid, warmed to room temperature and extracted with ethyl acetate. Theorganic layer was washed with water and saturated brine, dried, and thesolvent was removed in vacuo. The residue was purified by preparativeTLC to give 8.1 mg (quant) of 34. ¹H NMR (d6-aceton) δ: 1.23 (3H, t,J=7.3 HJz), 1.42 (12H, s), 3.04 (2H, q, J=7.5 Hz), 3.18 (2H, td, J=6.6,2.6 Hz), 3.31 (2H, t, J=6.6 Hz), 3.82 (4H, s), 6.99 (1H, brs), 7.08 (1H,t, J=2.6 Hz), 7.39 (2H, s).

Example 3

a. Compound 38

Using N-(4-methoxybenzyl)-1,2-isothiazolin-1,1-dioxide, 38 was preparedaccording to the synthesis of 28.

b. Methyl2-(5-(2-(4-Methoxy-benzyl)-1,1-dioxo-1,2-isothazolidin-5-ylidenemethyl)-3,3-dimethyl-2-oxo-2,3-dihydro-benzofuran-7-yl)-2-methyl-propionate(39)

To a solution of 38 (125 mg, 0.235 mmol) and Ph₃P (123 mg, 0.47 mmol) inTHF (3 ml), DEAD (0.056 ml, 0.353 mmol) was slowly added underice-cooling. After stirring for 1 hour, the reaction mixture wasconcentrated, and the residue was purified by silica gel chromatographyto yield 110 mg (92%) of 39. ¹H NMR (CDCl₃) δ: 1.26 (3H, t, J=7.2 Hz),1.50 (6H, s), 1.61 (6H, s), 3.02-3.18 (4H, m), 3.71 (3H, s), 3.81 (3H,s), 4.18 (2H, s), 6.87-6.90 (2H, m), 7.14 (1H, d, J=1.8 Hz), 7.26-7.33(4H, m).

c.2-(5-(1,1-Dioxo-1,2-isothiazolidin-5-ylidenemethyl)-2-hydroxy-3,3-dimethyl-2,3-dihydro-benzofuran-7-yl)-2-methyl-proplonicacid (41)

To a solution of 39 (63 mg, 0.160 mmol) in methanol (4 ml), 3N KOH (2ml) was added and the mixture was allowed to stand overnight at roomtemperature. The reaction mixture was concentrated, acidified withdiluted hydrochloric acid, and extracted with ethyl acetate. The organiclayer was washed with saturated brine, dried, and the solvent wasremoved in vacuo. The residue was added with dichloromethane (2 ml),cooled to −78° C., and dropwise and slowly added with DIBAL (0.93 M inn-hexane) (0.85 ml, 0.79 mmol). After 5 minutes, the mixture was addedwith diluted hydrochloric acid to quench the reaction and extracted withethyl acetate. The organic layer was washed with saturated brine, dried,and concentrated. The residue was purified by preparative TLC to give 38mg (63%) of 41. ¹H NMR (d6-aceton) δ: 1.29 (6H, br), 1.57 (6H, s), 3.24(2H, td, J=6.7, 2.5 Hz), 3.46 (2H, t, J=7.1 Hz), 5.67 (1H, s), 5.88(12H, t, J=7.7 Hz), 6.53 (1H, br), 7.13 (1H, t, J=2.5 Hz), 7.32 (1H, d,J=1.5 Hz), 7.38 (1H, d, J=1.5 Hz), 10.7 (1H, br).

Example 4

a. 3-tert-Butyl-2-methoxymethoxy-toluene (43)

NaH (60% in mineral oil, 4.5 g, 0.12 mol) is washed twice with hexane(50 ml) and added with THF (75 ml), and the mixture was cooled to below10° C. A solution of 2-tert-butyl-6-methylphenol 42 (16.4 g, 0.1 mol) inTHF (60 ml) was then dropwise and slowly added, and the mixture wasstirred for 10 minutes under the same conditions. The mixture was thendropwise added with MOMCl (9.1 ml, 0.12 mol), warmed to roomtemperature, and the reaction mixture was added with saturated aqueousammonium chloride (200 ml) and ethyl acetate (200 ml). The organic layerwas separated, washed sequentially with water (200 ml), 1.5N NaOH (200ml), water (100 ml), and saturated brine (200 ml), and dried. Thesolvent was removed in vacuo to give 20.5 g (98.6%) of 43 as colorlessoil.

b. 7-tert-Butyl-2(3H) -benzofuranone (44)

A solution of 43 (3.0 g, 14.4 mmol) and TMEDA (2.6 ml, 17.3 mmol) in THF(40 ml) was cooled to −50° C., and dropwise and slowly added withsec-BuLi (1.3 M in cyclohexane, 13.3 ml, 17.3 mmol). The reactionmixture was warmed to −30° C., and then cooled again to −60° C., and dryice was added to the mixture. The reaction mixture was warmed to roomtemperature over 1 hour, added with waster (50 ml) and ethyl acetate (50ml). The aqueous layer was separated, acidified with 2N HCl to pH 3-4,and extracted with ethyl acetate (100 ml). The organic layer was washedwith water (50 ml) and saturated brine (50 ml), and dried. The solventwas removed in vacuo. The residue was dissolved in toluene (50 ml),added with p-TsOH.H₂O (250 mg), and heated to reflux for 15 minutes. Thereaction mixture was cooled to room temperature and poured into anaqueous diluted sodium bicarbonate solution (60 ml). The organic layerwas separated and washed with saturated brine (60 ml). After drying, thesolvent was removed in vacuo, and the residue was purified by columnchromatography to yield 1.02 g (37%) of 44. ¹H NMR (CDCl₃) δ: 1.40 (9H,s, tBu), 3.71 (2H, s, CH₂), 7.02-7.27 (3H, m, 3×Ar—H).

c. 7-tert-Butyl-3,3-dimethyl-2(3H)-benzofuranone (45)

A solution of 44 (300 mg, 1.577 mmol), CH₃I (0.29 ml, 4.73 mmol) and18-Crown-6 (104 mg, 0.39 mmol) in THF (20 ml) was cooled to −50° C., andt-BuOK (389 mg, 3.47 mmol) was slowly added, and the mixture was stirredfor 1 hour. The mixture was warmed to room temperature and stirred foranother 1 hour. The reaction mixture was added with saturated aqueousammonium chloride (30 ml) and extracted with ethyl acetate (30 ml). Theorganic layer was washed with water (30 ml), and saturated brine (30ml), and dried. The solvent was removed in vacuo, and the residue waspurified by column chromatography to yield 241 mg (70%) of 45. ¹H NMR(CDCl₃) δ: 1.41 (9H, s, tBu), 1.50 (6H, s, 2×CH₃), 7.03-7.26 (3H, m,t3×Ar—H).

d. 2-tert-Butyl-6-(2-hydroxy-1,1-dimethylethyl)phenol (46)

A solution of 45 (235 mg, 1.077 mmol) in ether (5 ml) was dropwiseadded, with stirring and ice-cooling, to a suspension of LiAlH₄ (49 mg,1.29 mmol) in ether (5 ml). After washing a container of the solutionwith ether (5 ml), the washing was added to the mixture, which waswarmed to room temperature and stirred for 45 minutes. The reactionmixture was added with ethyl acetate (1 ml) and 3 ml of a mixture ofwater and saturated ammonium chloride (1:1), and stirred for 30 minutes.Resultant slurry was removed by filtration through celite, and thefiltrate was concentrated to obtain 253 mg (quant) of 46. ¹H NMR (CDCl₃)δ: 1.42 (9H, s, tBu), 1.44 (6H, s, 2×CH₃), 3.81 (2H, s, CH₂), 6.75-7.23(3H, m, 3×Ar—H), 9.20 (1H, br s, phonemic-OH).

e. 4-Bromo-2-tert-butyl-6-(2-hydroxy-1,1-dimethylethyl)phenol (47)

To a solution of 46 (1.297 g, 5.83 mmol) in dichloromethane (20 ml),Py.HBr₃ (1.86 g, 5.83 mmol) was added all at once, with stirring andice-cooling, and the mixture was stirred for 30 minutes. Water (30 ml)was added to the reaction mixture. The organic layer was separated andwashed with water (30 ml). After drying, the solvent was removed invacuo, and the residue was purified by column chromatography to yield1.346 g (76%) of 47. ¹H NMR (CDCl₃) δ: 1.39 (9H, s, tBu), 1.41 (6H, s,2×CH₃), 3.79 (2H, s, CH₂), 7.20 (1H, d, J=2.4 Hz, Ar—H), 7.27 (1H, d,J=2.4 Hz, Ar—H), 9.45 (1H, br s, phonemic-OH).

f. 4-Bromo-2-tert-butyl-6-(2-methoxymethoxy-1,1-dimetylethyl)phenylmethoxymethyl ether (48)

48 (930 mg, 56.9%) was prepared from 47 (1.264 g, 4.20 mmol), MOMCl(0.96 ml, 12.6 mmol) and NaH (402 mg, 10.5 mmol). ¹H NMR (CDCl₃) δ: 1.41(9H, s, tBu), 1.42 (6H, s, 2×CH₃), 3.31, 3.63 (each 3H, s, CH₃), 3.79(2H, s, CH₂), 4.59, 4.89 (each 2H, s, CH₂), 7.36 (1H, d, J=2.8 Hz,Ar—H), 7.38 (1H, d, J=2.8 Hz, Ar—H).

g.3-tert-Butyl-4-methoxymethoxy-5-(2-methoxymethoxy-1,1-dimethylethyl)benzaldehyde(49)

A solution of 48 (925 mg, 2.38 mmol) in THF (20 ml) was cooled to −50°C., and t-BuLi (1.7 M in pentane, 3.08 ml, 5.24 mmol) was dropwise addedto the solution while maintaining the temperature below −40° C. Aftercompletion of the addition, the mixture was stirred at this temperature,and DMF (0.12 ml) in THF (5 ml) was added. The mixture was warmed slowlyto room temperature. Aqueous solution of saturated ammonium chloride (50ml) was added, and the mixture was extracted with ethyl acetate (50 ml).The organic layer was washed with saturated brine (50 ml), and afterdryness, the solvent was removed in vacuo. The residue was purified bycolumn chromatography to yield 669 mg (83.1%) of 49 as colorless oil. ¹HNMR (CDCl₃) δ: 1.47 (9H, s, tBu), 1.48 (6H, s, 2×CH₃), 3.29, 3.66 (each3H, s, CH₃), 3.84 (2H, s, CH₂), 4.60, 4.95 (each 2H, s, CH₂), 7.81 (1H,d, J=2.2 Hz, Ar—H), 7.84 (1H, d, J=2.2 Hz, Ar—H), 9.92 (1H, s, CHO).

h.(E)-2-Ethyl-5-[3-tert-butyl-4-hydroxy-5-(2-hydroxy-1,1-dimethylethyl)benzylidene]-1,2-isothiazolidie-1,1-dioxide(51)

Aldol adduct 50 (868 mg, 91.3%) was obtained as colorless oil from 49(660 mg, 1.95 mmol) and N-Ethyl-1,2-isothiazolidine-1,1-dioxide (305.5mg, 2.05 mmol). To a solution of this adduct 50 (790 mg, 1.62 mmol) inmethanol (6 ml), about 10N HCl/MeOH (2 ml) was added, and the mixturewas stirred at room temperature for 30 minutes and at 50° C. for 30minutes. The reaction mixture was cooled to room temperature, andaqueous saturated sodium bicarbonate (50 ml) was added. The mixture wasextracted with ethyl acetate (50 ml), and the organic layer was washedwith saturated brine. After dryness, the solvent was removed in vacuo.Resultant residue was dissolved in toluene (35 ml). p-TsOH.H₂O (50 mg)was added, and the mixture was heated to reflux for 10 minutes. Thereaction mixture was cooled to room temperature, adsorbed to a smallamount of silica gel, which was eluted with ethyl acetate (150 ml). Thesolvent was removed in vacuo, and the resultant residue was purified bycolumn chromatography to yield 556 mg (90%) of 51 as colorless crystal.M.p. 152-155° C. IR (KBr) cm⁻¹: 3411, 3140, 2965, 1642, 1594, 1436,1291, 1266, 1143. ¹H NMR (CDCl₃) δ: 1.28 (3H, t, J=7.2 Hz, CH₃), 1.41(9H, s, tBu), 1.43 (6H, s, 2×CH₃), 2.95 (1H, t, J=3.8 Hz, alcoholic-OH),3.07-3.33 (6H, m, 2×CH₂, CH₂), 3.81 (2H, d, J=3.8 Hz, CH₂), 7.17 (1H, d,J=2.0 Hz, Ar—H), 7.23 (1H, t, J=2.0Hz, vinyl-H), 7.28 (1H, d, J=2.8 Hz,Ar—H), 9.92 (1H, s, phenolic-OH). Elementary analysis for C₂₀H₃₁NO₄S:Calcd: C, 62.96; H, 8.19; N, 3.67; S, 8.40. Found: C, 62.86; H, 8.19; N,3.70; S, 8.30.

i.(E)-5-(7-tert-Butyl-3,3-dimethyl-2,3-dihydrobenzofuran-5-ylmethylene)-2-ethyl-1,2-isothiazolidine-1,1-dioxide(52)

A mixture of 51 (125 mg, 0.328 mmol), MsCl (30 μl, 0.394 mmol),triethylamine (114 μl, 0.82 mmol) and dichloromethane (10 ml) wasstirred for 30 minutes under ice-cooling. The mixture was added withsaturated brine (30 ml) and extracted with dichloromethane (30 ml). Theorganic layer was separated, and after dryness, the solvent was removedin vacuo. The residue was purified by column chromatography to yield 81mg (68.1%) of 52. M.p. 104-106° C. IR (KBr) cm⁻¹: 3436, 2963, 2871,1642, 1602, 1453, 1288, 1151. ¹H NMR (CDCl₃) δ: 1.29 (3H, t, J=7.2 Hz,CH₃), 1.33 (6H, s, 2×CH₃), 1.35 (9H, s, tBu), 3.06-3.33 (6H, m, 2×CH₂,CH₂), 4.28 (2H, s, CH₂), 7.01 (1H, d, J=1.6 Hz, Ar—H), 7.15 (1H, d,J=1.6 Hz, Ar—H), 7.25 (1H, t, J=2.4 Hz, vinyl-H). Elementary analysisfor C₂₀H₂₉NO₃S: calcd: C, 66.08; H, 8.04; N, 3.85; S, 8.82. Found: C,66.14; H, 8.07; N, 3.82; S, 8.74.

The compound of the invention of formula Ia, as prepared in the abovePreparation 15, was evaluated in vivo experiments for its usefulness asan antirheumatic agent, as an osteoclast formation inhibiting agent, asa nitrogen monoxide production inhibiting agent, and as a transcriptionfactor NFκB suppressing agent.

Example 5 Antirheumatic Effect

1. Inhibitory Activity Against the Production of Cytokines

Cytokines are associated with production of proteases, collagenases,adhesion factors and the like, which are responsible for articulartissue destruction. Therefore, such articular tissue destruction couldbe suppressed by inhibition of cytokine production. One of thesecytokines, IL-6, was used to evaluate the inhibitory activity ofCompound Ia against the production of cytokine in the following manner.

[Method]

A neutrophil fraction from human peripheral blood was prepared, and 10μg/mL of LPS (lipopolysaccharide) and Compound Ia were added to thefraction, and the mixture was then incubated for 16-18 hours. IL-6 wasmeasured by RIA (radio immuno assay).

[Result]

Compound Ia inhibited the IL-6 production from neutrophil (IC₅₀: 20 μM)and inhibited the IL-6 production from synovial membrane cells of RA(rheumatic arthritis) patients (IC₅₀: 10 μM). Either of these inhibitoryactivities was greater than Tenidap, a comparative drug (Table 1).

TABLE 1 Inhibitory Activity Against IL-6 Production IC₅₀ (μM) synovialmembrane compound neutrophil cells from RA patient Compound Ia 20  10Tenidap 50 >50 Indomethacin nd >50 nd: not determined

2. Inhibitory Effect Against Rat Adjuvant Arthritis

A rat adjuvant arthritis model, which is commonly used in an in vivoevaluation for antirheumatic agents, was employed in this evaluation. Toprepare an arthritis model, heat-killed cells of Mycobacterium butyricum(Difco) were suspended in mineral oil, and a rat was intradermallyinjected with 0.05 mL of 1% solution of the suspension at the plantar ofleft hind leg.

(i) Inhibitory Effect Against Articular Swelling

[Method]

Using the above arthritis model, Compound Ia was administered to theanimal for 22 days after injection of the adjuvant, and the articularswelling was measured from time to time to evaluate the prophylacticinhibitory effect. On the other hand, the curative effect was evaluatedby administering Compound Ia to animals from Day 15 to Day 25 afterinjection of the adjuvant and measuring the articular swelling ininjected and non-injected legs. Inhibitory and curative effects wereexpressed by effective dose for 30% inhibition (ED₃₀).

[Result]

The results are graphically shown in FIG. 1 which shows the change involume of the adjuvant-injected or non-injected leg. Compound Iainhibited the articular swelling both in the adjuvant-injected andnon-injected legs, and dose-dependent and significant inhibitory effectwas observed particularly in the adjuvant-injected leg at a dose of 0.3mg/kg/day or more. On the other hand, as for the curative effect,dose-dependent and significant inhibitory effect was observed at a doseof 3 mg/kg/day or more.

ED₃₀ values for prophylactic and curative effects of Compound Ia and thecomparative drugs are shown in Table 2 below.

TABLE 2 Inhibitory Effect Against Adjuvant Articular Swelling ED₃₀(mg/kg/day) (95% confidence limits) compound prophylactic effectcurative effect Compound Ia 0.88(0.52-1.31) 2.6(1.3-6.4) Tenidap0.98(0.52-1.56) 4.9(2.4-23.0) Cyclosporin A 2.40(1.90-3.00) 4.7(1.9-7.6)Bucillamine >50 >50

(ii) Inhibitory Effect Against Articular Destruction

[Method]

The right and left hind legs of the arthritis rats used in the aboveexperiments were radiographed using a soft X-ray radiation and evaluatedby the X-ray observation score for the bone destruction in thearthrosis. The effects were numerically evaluated by grading thearticular destruction from 0 to 3 (0: normal; 1: low degree of articularand bone destruction; 2: medium degree of articular and bonedestruction, and neogenesis of cartilage; 3: strong articular and bonedestruction), and the efficacy in the drug-administered group wasdetermined in comparison with the vehicle-administered group accordingto Wilcoxon U-test (*p<0.05, **p<0.01) (FIG. 2).

[Result]

Compound Ia exhibited dose-dependent and significant inhibitory effectagainst the articular destruction at a dose of 3 mg/kg/day or more inthe adjuvant-injected leg and at a dose of 1 mg/kg/day or more in thenon-injected leg. The inhibitory effect of the comparative drug,Tenidap, was comparable to that of Compound Ia in the adjuvant-injectedleg, while the effect was lower than that of Compound Ia in thenon-injected leg, although the drug exhibited significant inhibitoryeffect at a dose of 3 mg/kg/day. On the other hand, Cyclosporin Aexhibited significant inhibitory effect at a dose of 10 mg/kg/day bothin the adjuvant-injected and non-injected legs (FIG. 3).

(iii) Repressive Effect on the IL-6 Level in Articular Tissue

[Method]

Another adjuvant arthritis model animal was prepared apart from theabove experiments, and a compound was orally administered to the animalfor 18 days after the adjuvant injection. The non-injected leg was thenamputated under anesthetization, epidermis of the amputated leg wasablated, the leg was frozen in liquid nitrogen, and the arthrosis partwas crushed by a crushing machine. To a measured quantity of the crushedarthrosis, 3 mL of ice-cold PBS was added and homogenized with ahomogenizer under ice-cooling. The resultant homogenate was centrifugedat 4° C. for 20 minutes at 3,000 rpm to collect the supernatant, whichwas then passed through a 0.45 μm filter to prepare a sample. The sampleand MH60 BSF2 cells (derived from mouse, require IL-6) were incubated ona 96-well plate under defined conditions, and the absorbance wasmeasured at two wave lengths of 540 nm and 690 nm. The IL-6 level in thesample was calculated based on the standard curve prepared using arecombinant human IL-6. The results were shown using mean±standard errorof the data of seven animals, and significance test was performedaccording to Dunnett-t test (##p<0.01: vs. normal group; **p<0.01: vs.vehicle group).

[Result]

The IL-6 level in the articular tissue of the adjuvant arthritis rats(Veh.) was 44.2 μg/g tissue, indicating remarkable increase of IL-6level compared to that of normal rats (Nor.). Compound Ia exhibiteddose-dependent and significant inhibitory effect at a dose of 3mg/kg/day or more against the increase of the IL-6 level in thearticular tissue of pathologic animals. Tenidap and Bucillamine alsoexhibited significant inhibitory effect at a dose of 10 mg/kg/day, andtheir efficacies were comparable to that of Compound Ia. Cyclosporin A(CsA) at a dose of 20 mg/kg/day almost completely inhibited the increaseof the IL-6 level (FIG. 4).

3. Inhibitory Effect Against Bone Destruction in

Spontaneous Rheumatic Model Animal NZB/KN Mouse

[Method]

To male NZB/KN mice, in which a human RA-like pathology arisesspontaneously due to their immunologically abnormal background, wasorally administered Compound Ia once in a day (five times a week) forsix months beginning at two-month-age. The mice which reached six monthold (some were eight month old) were full-length radiographed underanesthetization and predetermined conditions using a soft X-raygenerator (OM-55, Omic). According to the method of Nakamura, et. al.[Arthritis. Rheum., 34, 171-179 (1991)], the number of the lesionsobserved in the bones and articular regions in the whole body werecounted on the basis of the soft X-ray radiographed films, and the totalnumber of the observed lesions were defined as the number of the bonedisorders in the animal. The results were shown using mean±standarddeviation, and significance test was performed according to Dunnett-ttest (*p<0.05, **p<0.01: vs. vehicle group).

[Result]

Compound Ia exhibited significant improving effect on the bone-articularlesions at a dose of 10 mg/kg/day or more. As for the comparative drugs,Tenidap (Ten) and Cyclosporin A (CsA), which were each administered at adose of 30 mg/kg/day, Cyclosporin A exhibited significant inhibitoryeffect while Tenidap was not effective. On the other hand, Indomethacinat a dose of 2 mg/kg/day exhibited no inhibitory effect but significantenhancing effect on the bone destruction (FIG. 5).

4. Effect on MRL/l Mouse Blood Acute Phase Protein (Serum AmyloidProtein: SAP)

[Method]

To MRL/l male mice known as a lupus mouse in which a human RA-likepathology spontaneously arises, Compound Ia was orally administered fivetimes a week (twice a week for Predonisolone) for 13 weeks. After thelast administration of the compound, blood was collected underanesthetization, and the serum was used for SAP measurement. SAP hasbeen used as a marker for rheumatism. A 96-well plate was coated withrabbit anti-mouse SAP serum, and the serum to be tested was then addedto the plate and incubated for 2 hours. After addition of sheepanti-mouse SAP serum, a rabbit anti-sheep IgG antibody labeled withalkaline phosphatase and then a substrate were added to the plate, andOD of developed color was measured at 405 nm. The results were shownusing mean±standard error, and significance test of the results wasperformed according to Dunnett-t test (*p<0.05, **p<0.01: vs. vehiclegroup).

[Result]

The groups which received Compound Ia at a dose of 2-50 mg/kg/dayexhibited repressive tendency in a dose-dependent manner, and the doseof 50 mg/kg/day gave significant results. Significant inhibitory effectswere also observed in the groups which received comparative drugs,Lobenzarit (Lob.) at a dose of 25 mg/kg/day and Predonisolone (Pred.) ata dose of 10 mg/kg/day. However, the groups which received Tenidap(Ten.) at a dose of 10 mg/kg/day, Indomethacin (Ind.) at a dose of 1mg/kg/day and Bucillamine (Buc.) at a dose of 10 or 25 mg/kg/day gave nosignificant results although they exhibited suppressive tendency (FIG.6).

Example 6 Inhibitory Effect Against Osteoclast Formation

Inhibitory Effect against enhanced formation of Osteoclast in MouseMyeloblast

[Method]

It has been known that the enhanced formation of osteoclast is inducedin the mouse myeloblast culture by addition of IL-1β and such effect isderived from PGE₂ production through COX-2 induction. In order todetermine inhibitory effect of the compounds against such effect,myeloblasts were collected from a ddY male mice, plated in a 48-wellplate at 1×10⁶/well and incubated. After culturing for six days in anα-MEM medium containing 10% fetal bovine serum, to which 2.5 ng/mL ofhuman IL-1β and a predetermined concentration of the compounds wereadded, PGE₂ level in the supernatant was measured by RIA kit. On theother hand, cells adhering on the plate were immobilized, and anactivity staining of tartaric acid resistant acid phosphatase (TRAP),which is a marker enzyme for osteoclast, was conducted, whereby thenumber of TRAP positive cells having multinucleate (N>3) per well wascounted as osteoclast.

[Result]

Test compounds were added to the above culture and assayed for theirinhibitory activity against the PGE₂ production due to IL-1β stimulationand the enhanced formation of osteoclast. The results were shown interms of IC₅₀ (Table 3). Compound Ia exhibited stronger inhibition ascompared with Tenidap and Indomethacin. The inhibition of Compound Iawas greater than that ofN-[2-(cyclohexyloxy)-4-nitrophenyl]-methanesulfoneamide, a selectiveCOX-2 inhibitor.

TABLE 3 Inhibitory Activity Against Enhanced Formation of Osteoclast andPGE₂ Production IC₅₀ (μM) Inhibition of Inhibition of Osteoclast PGE₂compound Formation Production Compound Ia 0.0013 0.00021 Tenidap 0.0630.0017 Indomethacin 0.032 0.0064 N-[2-(cyclohexyloxy)nitro- 0.024 0.0042phenyl)methanesulfoneamide

Example 7 Inhibitory Effect Against Nitrogen Monoxide Production

1. Inhibitory Activity against NO Production from Activated Macrophage(MΦ)

[Method]

To BALB/c male mice was intraperitoneally administered 2 mL of athioglycolate solution, and induced activated MΦ were collected afterfour days and used as NO producing cells. The cells were adjusted to2×10⁶cells/mL and plated onto a 96-well microtiter plate at2×10⁵cells/100 μL/well. 100 μL of 1 μg/mL of Compound Ia and LPS(lipopolysaccharide) were added thereto and incubated at 37° C. under 5%CO₂ for 24 hours. 75 μL of the supernatant obtained after incubation and75 μL of Griess reagent were incubated, and NO²⁻ in the form of sodiumnitrite was measured to determine the amount of NO in the supernatant.

[Result]

Compound Ia at 0.8-12.5 μM inhibited NO production in a dose-dependentmanner in the above assay system. The inhibitory activity of Compound Iaagainst NO production was greater than those of the comparative agents,Tenidap and Indomethacin (Table 4).

TABLE 4 Inhibitory Activity Against NO Production In Mouse ActivatedMacrophage NO²⁻ Production (μM/2 × 10⁵ cells) Medium <0.1 Medium + LPS28.3 ± 2.29 μM Compound Ia Tenidap Indomethacin 0.8 27.3 ± 0.94 31.5 ±1.05 29.5 ± 0.68 3.1 12.3 ± 2.50** 33.1 ± 0.75 27.9 ± 1.91 12.5 <1.0***36.2 ± 0.91* 23.9 ± 2.49 50 <1.0*** 19.7 ± 0.96** 14.9 ± 3.02** 100<1.0***  3.3 ± 1.99***  3.0 ± 2.41*** IC₅₀ (μM)  3.0 75.0 60.0 Values inthe table are mean ± standard error of independent experiments. *p <0.05, **p < 0.01, ***p < 0.001: Student t-test

2. Suppressive Effect on Blood NO Level

[Method]

An adjuvant arthritis model was prepared in the same manner as above,and the compounds were orally administered to the animals for 28 daysbeginning on the day before the adjuvant injection. Blood was thencollected under anesthetization and the serum was ultracentrifuged. Thesupernatant was ultrafiltrated to prepare a sample for NOx(nitrite+nitrate) measurement. Serum NOx level was determined by themeasurement of optical density at 550 nm using the improvedabsorptiometry by Griess [Tracy, W. R. Tse, J., and Carter, G., (1995),J. Pharmacol. Exp. Ther., 272, 1011-1015]. The results were shown usingmean±standard error of the data of eight samples, and significance testwas performed according to Dunnett's t-test (##p<0.01: vs. normal group;*p<0.05, **p<0.01: vs. vehicle group).

[Result]

Serum NOx level in the arthritis-maturing phase on 28th day afteradjuvant administration was increased by about three times as comparedwith the normal level. Compound Ia at a dose of 3 mg/kg/day or moreexhibited the dose-dependent and significant inhibitory effect on theincrease of blood NOx level in this pathological animal. As for thecomparative drug, Tenidap, significant effect was observed at 10mg/kg/day, while mere inhibitory tendency was observed at 30 mg/kg/day.Thus, the inhibitory effect of Tenidap was lower than that of CompoundIa. Indomethacin (Ind) at 1 mg/kg/day and Dexamethazon (Dex) at 0.3mg/kg/day also exhibited significant inhibitory effect (FIG. 7).

Example 8 Suppressive Effect on Transcription Factor NFκB

Suppressive activity on Transcription Factor NFκB

[Method]

A recombinant plasmid (pGV-B.HIV-1LTR), in which HIV-1 (AIDS) viruspromoter LTR containing two NFκB binding sites is ligated to upstream offirefly luciferase gene, was constructed, and the DNA was purified. Thisplasmid DNA was introduced by electroporation into a Jurkat cell, ahuman T cell leukemic cell line, and the cell was cultured for 48 hours.Subsequently, phytohemagglutinin (PHA, 0,5%), phorbol ester (TPA(12-O-tetradecanoylphorbol 13-acetate), 25 ng/mL), and a predeterminedconcentration of the test compound were added thereto to allow to reactfor 2 hours, and the cells were harvested, and the luciferase activityin the cellular extract was measured.

[Result]

Compound Ia inhibited luciferase activity at 5-50 μM in a dose-dependentmanner (Table 5, Experiment-1). The comparative drugs, Tenidap andLobenzarit, inhibited the activity at 20-50 μM and at 500 μM,respectively, while their efficacies were lower than that of CompoundIa. No inhibitory effect was observed for Bucillamine even at , 500 μM(Table 5, Experiment-1 and Experiment-2).

TABLE 5 Suppressive Activity on Transcription Factor NFκB luciferaseactivity (%) mean ± μM N standard error Experiment-1 control 2  26.9 ±1.5 TPA-PHA 4 100.0 ± 2.5 Compound Ia 5 4  82.2 ± 8.3 10 4  79.6 ± 5.420 4  54.6 ± 4.3 50 4  53.5 ± 5.9 Tenidap 5 4  94.0 ± 14.0 10 4  94.4 ±7.2 20 3  75.6 ± 19.1 50 4  69.3 ± 17.1 Experiment-2 control 2  44.1 ±9.4 TPA-PHA 2 100.0 ± 4.6 Compound Ia 20 2  86.4 ± 1.9 50 1  75.1 100 2 61.1 ± 3.9 Lobenzarit 20 2 112.6 ± 1.9 100 2 100.6 ± 2.7 500 2  70.6 ±4.9 Bucillamine 20 1  96.0 100 2  96.1 ± 6.2 500 2 101.0 ± 0.8 Values inthe table are mean ± standard error of independent experiments in termsof relative luciferase activity.

Example 9

Formulations 1) Granules Compound of the invention  20 mg Lactose 250 mgCorn starch 115 mg Hydroxypropylcellulose 115 mg

Above materials are Granulated in a Conventional Wet process to obtainGranules.

EFFECT OF THE INVENTION

The compounds of the present invention have an inhibitory effect againstarticular bone destruction and the like in a rheumatic model animal, andyet the inhibitory effect against the production of JL-6 and acute phaseproteins such as serum amyloid. Accordingly, the compounds of theinvention are useful as an antirheumatic agent for prevention and/ortreatment of rheumatoid arthritis.

Also, since the compounds of the present invention have an inhibitoryeffect against the enhanced formation of osteoclast in mousemyeloblasts, they are useful as an preventing and/or treating agent forbone metabolic disorders such as rheumatoid arthritis, osteoarthritisand osteopprosis and the like.

Further, the compounds of the present invention are useful forprevention and/or treatment of rheumatoid arthritis and osteoarthritissince they exhibit an inhibitory effect against NO production inmacrophage which has received an inflammatory stimulus and NO productionin an inflammatory pathological model animal. Due to inhibitory effectagainst NO production, the compounds of the invention would be usefulfor prevention and/or treatment of atherosclerosis, ischemic heart andbrain disease, Alzheimer disease, diabetes, endotoxin shock, sepsis,ulcerative colitis and the like.

Furthermore, since the compounds of the invention exhibit a suppressiveeffect on NFκB in human cells, they are useful for prevention and/ortreatment of chronic articular rheumatism. Based on the suppressiveeffect on NFκB, the compounds of the invention would also be useful forsystemic lupus erythematosus, Behcet's disease, ulcerative colitis,atherosclerosis, endotoxin shock, sepsis, cytomegalovirus pneumonia,adenovirus cold, AIDS and the like.

What is claimed is:
 1. A method for inhibition of osteoclast formationwhich comprises administering to mammals in need thereof an effectiveamount of the compound of formula I:

wherein R¹ is hydrogen, lower alkyl, lower alkoxy, hydroxy lower alkyl,or carboxy lower alkyl and R⁴ is hydroxy; or R¹ and R⁴ taken togetherform —CR⁵R⁶—(CH₂)_(m)—O— or —CR⁵R⁶—(CH₂)_(p)CH(OH)—O— wherein m is aninteger of 1 to 3; p is an integer of 0 to 2; and R⁵ and R⁶ are eachindependently hydrogen, lower alkyl, lower alkoxy, or hydroxy loweralkyl; R² is hydrogen, lower alkyl, lower alkoxy, hydroxy lower alkyl,or carboxy lower alkyl; and R³ is hydrogen, lower alkyl, cycloalkyl,lower alkoxy, arylalkyloxy, heteroarylalkyloxy, lower alkylcarbonyl,arylcarbonyl, substituted or unsubstituted carbamoyl or a grouprepresented by the formula: —(CH₂)_(n)—Q wherein Q is hydroxy,substituted or unsubstituted amino, aryl, heteroaryl, hydroxycarbonyl orlower alkyloxycarbonyl; and n is an integer of 0 to 3 or apharmaceutically acceptable salt or hydrate thereof.
 2. The method ofclaim 1, wherein R¹ and R² are both t-butyl, and R⁴ is hydroxy.
 3. Themethod of claim 1, wherein R¹ and R² are both t-butyl, R³ is loweralkyl, and R⁴ is hydroxy.
 4. A method for prevention and/or treatment ofrheumatoid arthritis which comprises administering to mammals in needthereof an effective amount of the compound of formula I:

wherein R¹ is hydrogen, lower alky, lower alkoxy, hydroxy lower alkyl,or carboxy lower alkyl and R⁴ is hydroxy; or R¹ and R⁴ taken togetherform —CR⁵R⁶—(CH₂)_(m)—O— or —CR⁵R⁶—(CH₂)_(p)CH(OH)—O— wherein m is aninteger of 1 to 3; p is an integer of 0 to 2; and R⁵ and R⁶ are eachindependently hydrogen, lower alkyl, lower alkoxy, or hydroxy loweralkyl; R² is hydrogen, lower alkyl, lower alkoxy, hydroxy lower alkyl,or carboxy lower alkyl; and R³ is hydrogen, lower alkyl, cycloalkyl,lower alkoxy, arylalkyloxy, heteroarylalkyloxy, lower alkylcarbonyl,arylcarbonyl, substituted or unsubstituted carbamoyl or a grouprepresented by the formula: —(CH₂)_(n)—Q wherein Q is hydroxy,substituted or unsubstituted amino, aryl, heteroaryl, hydroxycarbonyl orlower alkyloxycarbonyl; and n is an integer of 0 to 3 or apharmaceutically acceptable salt or hydrate thereof.
 5. The method ofclaim 4, wherein R¹ and R² are both t-butyl, and R⁴ is hydroxy.
 6. Themethod of claim 4, wherein R¹ and R² are both t-butyl, R³ is loweralkyl, and R⁴ is hydroxy.
 7. A method for inhibition of nitrogenmonoxide production which comprises administering to mammals in needthereof an effective amount of the compound of formula I:

wherein R¹ is hydrogen, lower alkyl, lower alkoxy, hydroxy lower alkyl,or carboxy lower alkyl and R⁴ is hydroxy; or R¹ and R⁴ taken togetherform —CR⁵R⁶—(CH₂)_(m)—O— or —CR⁵R⁶—(CH₂)_(p)CH(OH)—O— wherein m is aninteger of 1 to 3; p is an integer of 0 to 2; and R⁵ and R¹ are eachindependently hydrogen, lower alkyl, lower alkoxy, or hydroxy loweralkyl; R² is hydrogen, lower alkyl, lower alkoxy, hydroxy lower alkyl,or carboxy lower alkyl; and R³ is hydrogen, lower alkyl, cycloalkyl,lower alkoxy, arylalkyloxy, heteroarylalkyloxy, lower alkylcarbonyl,arylcarbonyl, substituted or unsubstituted carbamoyl or a grouprepresented by the formula: —(CH₂)_(n)—Q wherein Q is hydroxy,substituted or unsubstituted amino, aryl, heteroaryl, hydroxycarbonyl orlower alkyloxycarbonyl; and n is an integer of 0 to 3 or apharmaceutically acceptable salt or hydrate thereof.
 8. The method ofclaim 7, wherein R¹ and R² are both t-butyl, and R⁴ is hydroxy.
 9. Themethod of claim 7, wherein R¹ and R² are both t-butyl, R³ is loweralkyl, and R⁴ is hydroxy.
 10. A method for suppression of transcriptionfactor NFκB which comprises administering to mammals in need thereof aneffective amount of the compound of formula I:

wherein R¹ is hydrogen, lower alkyl, lower alkoxy, hydroxy lower alkyl,or carboxy lower alkyl and R⁴ is hydroxy; or R¹ and R⁴ taken togetherform —CR⁵R⁶—(CH₂)_(m)—O— or —CR⁵R⁶—(CH₂)_(p)CH(OH)—O— wherein m is aninteger of 1 to 3; p is an integer of 0 to 2; and R⁵ and R⁶ are eachindependently hydrogen, lower alkyl, lower alkoxy, or hydroxy loweralkyl; R² is hydrogen, lower alkyl, lower alkoxy, hydroxy lower alkyl,or carboxy lower alkyl; and R³ is hydrogen, lower alkyl, cycloalkyl,lower alkoxy, arylalkyloxy, heteroarylalkyloxy, lower alkylcarbonyl,arylcarbonyl, substituted or unsubstituted carbamoyl or a grouprepresented by the formula: —(CH₂)_(n)—Q wherein Q is hydroxy,substituted or unsubstituted amino, aryl, heteroaryl, hydroxycarbonyl orlower alkyloxycarbonyl; and n is an integer of 0 to 3 or apharmaceutically acceptable salt or hydrate thereof.
 11. The method ofclaim 10, wherein R¹ and R² are both t-butyl, and R⁴ is hydroxy.
 12. Themethod of claim 10, wherein R¹ and R² are both t-butyl, R³ is loweralkyl, and R⁴ is hydroxy.